OSAC 2022-S-0023, Standard Practice for the Forensic

Analysis of Explosives By Polarized Light Microscopy

OSAC 2022-S-0023
Standard Practice for the
Forensic Analysis of
Explosives By Polarized Light
Microscopy

Ignitable Liquids, Explosives, and Gunshot Residue Subcommittee

Chemistry: Trace Evidence Scientific Area Committee (SAC)
Organization of Scientific Area Committees (OSAC) for Forensic Science
 OSAC 2022-S-0023, Standard Practice for the Forensic
Analysis of Explosives By Polarized Light Microscopy

Draft OSAC Proposed Standard

OSAC 2022-S-0023
Standard Practice for the Forensic
Analysis of Explosives By Polarized
Light Microscopy
Prepared by
Ignitable Liquids, Explosives, and Gunshot Residue Subcommittee
Version: 1.0
February 2021

Disclaimer:
This OSAC Proposed Standard was written by the (subcommittee) of the Organization of Scientific Area
Committees (OSAC) for Forensic Science following a process that includes an open comment period.
This Proposed Standard will be submitted to a standards developing organization and is subject to
change.
There may be references in an OSAC Proposed Standard to other publications under development by
OSAC. The information in the Proposed Standard, and underlying concepts and methodologies, may be
used by the forensic-science community before the completion of such companion publications.
Any identification of commercial equipment, instruments, or materials in the Proposed Standard is not a
recommendation or endorsement by the U.S. Government and does not imply that the equipment,
instruments, or materials are necessarily the best available for the purpose.
To be placed on the OSAC Registry, certain types of standards first must be reviewed by a Scientific and
Technical Review Panel (STRP). The STRP process is vital to OSAC’s mission of generating and
recognizing scientifically sound standards for producing and interpreting forensic science results. The
STRP shall provide critical and knowledgeable reviews of draft standards or of proposed revisions of
standards previously published by standards developing organizations (SDOs) to ensure that the
published methods that practitioners employ are scientifically valid, and the resulting claims are
trustworthy.
The STRP panel will consist of an independent and diverse panel, including subject matter experts,
human factors scientists, quality assurance personnel, and legal experts, which will be tasked with
evaluating the proposed standard based on a comprehensive list of science-based criteria.

2
 OSAC 2022-S-0023, Standard Practice for the Forensic
Analysis of Explosives By Polarized Light Microscopy

For more information about this important process, please visit our website
at: https://www.nist.gov/topics/organization-scientific-area-committees-forensic-science/scientific-
technical-review-panels.

3
 OSAC 2022-S-0023, Standard Practice for the Forensic
Analysis of Explosives By Polarized Light Microscopy
1 Standard Practice for the Forensic Analysis of Explosives By Polarized Light Microscopy
2 __________________________________________________________________________
3
4 1. Scope
5
6 1.1 This practice addresses the use of polarized light microscopy (PLM) to identify explosive -related
7 compounds from intact explosives and post-blast residues containing unconsumed explosive compounds,
8 and to isolate them for further analysis.
9 1.2 This practice is intended for use by competent forensic science practitioners with the requisite formal
10 education, discipline-specific training (see Practice E2917), and demonstrated proficiency to perform
11 forensic casework (refer to the T/SWGFEX Suggested Guide for Explosives Analysis Training).
12 1.3 This practice does not purport to address all of safety concerns, if any, associated with its use. It is the
13 responsibility of the user of this standard to establish appropriate safety and health practices and
14 determine the applicability of regulatory limitations prior to use.
15 2. Referenced Documents
16
17 2.1 ASTM Standards
18 E620 Practice for Reporting Opinions of Scientific or Technical Experts
19 E860 Practice for Examining and Preparing Items that are or may become Involved in Criminal or
20 Civil Litigation
21 E1732 Terminology Related to Forensic Science
22 E2917 Practice for Forensic Science Practitioner Training, Continuing Education, and Professional
23 Development Program
24 WK56998 Terminology Relating to the Examination of Explosives
25 WK67862 Practice for Establishing an Examination Scheme for Intact Explosives
26
27 2.2 Other Resources
28 Technical/Scientific Working Group for Fire and Explosion Analysis (T/SWGFEX) Suggested Guide for
29 Explosive Analysis Training
30 (https://www.nist.gov/system/files/documents/2018/09/21/twgfex_suggest_guide_for_explosive_analysis
31 _training.pdf)
32
33 3. Terminology
34
35 3.1 Definitions - For definitions of terms used in this guide other than those listed in 3.2, see E1732 and
36 WK56998.
37
38 3.2 Definitions of Terms Specific to This Standard:
39
40 alpha (α) - symbol representing the lowest of the three principal refractive indices of a biaxial crystal.
41
42 anomalous interference colors, n – atypical polarization colors which occur due to variation with
43 wavelength of the refractive index; very few substances display this characteristic.
44
45 Becke line, n – a halo observed near the boundary of a transparent particle when mounted in a medium
46 that differs from its refractive index.
47
 OSAC 2022-S-0023, Standard Practice for the Forensic
Analysis of Explosives By Polarized Light Microscopy
48 Becke line method, n - method for determining the refractive index of a transparent particle relative to its
49 mountant by noting the direction in which the Becke line moves when the focus is changed.
50
51 Discussion--The Becke line always moves toward the higher refractive index medium (particle or
52 mountant) when the focus is raised (stage is lowered) ,and towards the lower refractive index medium
53 when the focus is lowered (stage is raised). At the point where the index of the particle matches the
54 index of the mounting medium, the Becke line is no longer visible. The Becke line is generally viewed at
55 a wavelength of 589 nm (the D line of Sodium [nD]).
56
57 beta (β) - symbol representing the intermediate principal refractive index of a biaxial crystal.
58
59 biaxial, adj - an anisotropic crystal in the orthorhombic, monoclinic or triclinic system with three
60 principal refractive index directions (α, β, γ) and two optic axes that are isotropic.
61
62 birefringence, n - the numerical difference between the maximum and minimum refractive indices of
63 anisotropic substances: ε − ω for uniaxial crystals and γ − α for biaxial crystals
64
65 crystal system, n - crystals are classified according to their crystallographic axes length and the angles
66 between them. There are six crystal systems: cubic, tetragonal, hexagonal, orthorhombic, monoclinic, and
67 triclinic. All crystal systems are anisotropic except for the cubic system, which is isotropic.
68
69 epsilon (ε) - any vibration direction in the plane of the c axis for uniaxial crystals.
70
71 euhedral, adj - well-formed crystals bounded by plane faces
72
73 eutectic, n - the lowest melting mixture of two or more substances miscible in the melt state.
74
75 extinction, n - condition when an anisotropic substance appears dark when observed between crossed
76 polarizers and occurs when the vibration directions of the specimen are parallel to the vibration directions
77 of the polarizer and analyzer.
78
79 extinction, oblique, n - a type of extinction in which the vibration directions are oblique to the long
80 direction of the crystal
81
82 extinction, parallel, n - a type of extinction in which the vibration directions are parallel and perpendicular
83 to the long direction of the crystal
84
85 extinction, symmetrical, n - a type of extinction in which the vibration directions bisect a prominent
86 interfacial angle of the crystal
87
88 gamma (γ) - symbol representing the highest of the three principal refractive indices of a biaxial crystal.
89
90 interference figure, n – pattern observed during conoscopic examination of an anisotropic material, which
91 consists of a combination of extinction positions and interference colors corresponding to the full cone of
92 directions by which the sample is illuminated.
93
94 fusion methods, n— A process in which compounds are heated on a microscope slide and observed via
95 PLM during heating, melting and, upon cooling, recrystallization of the melt.
96 Discussion—The procedure frequently employs a temperature controlled hot stage capable of at least
97 300O Celsius placed on the stage of a polarizing light microscope.

2
 OSAC 2022-S-0023, Standard Practice for the Forensic
Analysis of Explosives By Polarized Light Microscopy
98 omega (ω) - Any vibration direction in the plane of the a axes for uniaxial crystals
99 optic axial angle (2V), n - the acute angle between two optic axes of a biaxial crystal.
100
101 optic axis, n - a direction of isotropic light propagation in an anisotropic crystal. Uniaxial crystals have
102 one optic axis; biaxial crystals have two optic axes.
103
104 optic sign, n - determined by the relationship of the refractive indices of a material; for uniaxial crystals:
105 if ε > ω, the crystal is positive (+); if ω > ε, the crystal is negative (-); for biaxial crystals: if γ-β > β-α, the
106 crystal is positive (+); if γ-β < β- α, the crystal is negative (-)
107
108 polymorphism, n - crystallization into two or more chemically identical but crystallographically distinct
109 forms.
110
111 relief, n - contrast between a particle or crystal and its media due to the difference between their refractive
112 indices. The greater the numerical difference in refractive indices, the greater the relief.
113
114 subhedral, adj - crystals with imperfectly developed faces
115
116 uniaxial, adj - an anisotropic crystal in the tetragonal or hexagonal system having one optic axis (isotropic
117 direction) and either two (tetragonal) or three (hexagonal) directions which are alike and perpendicular to
118 the direction of the optic axis.
119
120 4. Summary of Practice
121
122 4.1 Solid chemical components present in explosives are analyzed using a polarized light microscope
123 (PLM). The optical crystallographic properties such as refractive index, birefringence, and morphology
124 are determined. Determination of these properties, which are different for the various explosive
125 components, can aid in the identification of explosives and explosive residues submitted to the forensic
126 laboratory.
127
128 4.2 Optical crystallographic properties of an unknown component can be determined by mounting in a
129 refractive index liquid, by recrystallizing the component from a small drop of solvent, or recrystallizing
130 from its melt, i.e., recrystallization occurring when a melted compound is cooled. Recrystallized
131 components can be isolated for further analysis using other analytical techniques such as Fourier
132 transform infrared spectroscopy (FTIR), Raman spectroscopy, or scanning electron microscopy-energy
133 dispersive electron spectroscopy (SEM-EDS).
134
135 4.3 Water-soluble components present in explosives, such as inorganic salts used as oxidizers, are
136 dissolved in water and tested using reagents to form characteristic microcrystals. Microcrystal tests can
137 aid in the determination of anions and cations present in oxidizers and other components of explosives.
138
139 4.4 The optical crystallographic properties of a compound and microchemical testing for its ions are
140 independent techniques that provide complementary information. A combination of tests that include the
141 optical crystallographic characterization of a compound and microchemical tests to confirm the anion and
142 cation are used to identify oxidizers and other salts present in explosives.
143
144 4.5 A combination of tests that include optical crystallographic characterization or microchemical tests
145 are used in conjunction with other independent techniques such as FTIR or SEM-EDS to identify
146 chemical components present in explosives. Refer to ASTM WK67892 for the requirements for the

3
 OSAC 2022-S-0023, Standard Practice for the Forensic
Analysis of Explosives By Polarized Light Microscopy
147 identification of commonly encountered explosives and chemical components present in explosive
148 materials.
149
150 5. Significance and Use
151
152 5.1 This practice is designed to assist the analyst using polarized light microscopy for the identification of
153 explosives.
154
155 5.2 Intact explosives and post-blast residues that contain unconsumed explosive components are suitable
156 for this practice.
157
158 5.3 Polarized light microscopy is not appropriate for the examination of smokeless powder.
159
160 5.4 Some of the techniques described in this practice allow for the recovery of the specimen, but others
161 such as microcrystal tests are destructive.
162
163 6. Apparatus
164
165 6.1 Polarized light microscope - A compound microscope with a rotating stage, polarizing filters,
166 condenser and compensator slot. Most commercially available polarized light microscopes should be
167 adequate for the forensic analysis of explosives.
168 6.2 Objectives - Strain-free 10X, 20X and 40X. A 40X objective with a numerical aperture of 0.65 or
169 higher is recommended to provide the widest view of the back focal plane for conoscopic observations.
170 6.3 Compensator - A full wave compensator for use with the microscope.
171 6.2 Hot stage - a temperature controlled hot stage for use with a microscope with a range of
172 approximately 30oC to 300oC.
173 6.5 The best practice for documenting some microscopical observations, such as crystal formations, is to
174 take a digital photograph. It is advised that the minimum equipment required also has the capability of
175 digital photography.
176 6.6 Alcohol lamp
177 6.7 Glass ring with a diameter of approximately 10-12 mm and 5-10 mm in height
178
179 7. Reagents and Materials
180
181 7.1 Microscope slides and cover slips
182 7.2 Refractive index oils
183 7.3 Reagents for microcrystal tests: chloroplatinic acid, zinc/uranyl acetate, ammonium molybdate,
184 squaric acid, potassium iodide, nitron sulfate, strychnine sulfate, methylene blue
185 7.4 Reference materials, including reagent grade chemicals and explosive materials, can be obtained from
186 commercial and retail sources or directly from the distributor or manufacturer.
187 7.5 Distilled or deionized water
188 7.6 Toluene or methylene chloride, reagent grade
189 7.7 Sodium hydroxide, reagent grade
190 7.8 Glass rod with a tapering tip of approximately 1 mm or less in diameter
191 7.9 Glass ring with an approximate diameter of 10-12 mm
192 7.10 Tungsten needles
193 7.11 589 nm light filter
194
195 8. Initial Microscopical Examination

4
 OSAC 2022-S-0023, Standard Practice for the Forensic
Analysis of Explosives By Polarized Light Microscopy
196 8.1. If preliminary examinations, such as stereomicroscopical observations, ignition susceptibility test
197 (IST), and chemical spot tests indicate that the specimen is an explosive mixture or a component thereof,
198 1-3 particles or grains (approximately 1 mg or less) are mounted on a glass microscope slide with a cover
199 slip and dispersed in a refractive index oil. Any refractive index oil in the 1.400 to 1.660 range can be
200 used, however, if preliminary examinations indicate a particular explosive material, e.g., black powder,
201 ammonium nitrate-fuel oil (ANFO) or Composition C-4, the examiner can select a refractive index oil
202 that will provide characteristic optical crystallographic data based upon Tables 1 and 2.
203 8.2 Examine the mounted sample using a polarized light microscope using a 10X objective. Increase
204 magnification as needed.
205 8.3 Charcoal, if present, will appear as irregularly shaped, black and opaque particles, and as brown flakes
206 often with cellular features such as pits or cell walls.
207 8.4 Most common explosives and oxidizers are colorless when viewed with a transmitted light
208 microscope. Using crossed polarizers, determine if the crystals are isotropic or anisotropic. Assess the
209 interference colors to determine if the birefringence is low (less than 0.01), moderate (0.01-0.10) or high
210 (0.10 and above).
211 8.5 Determine relative refractive indices of crystals present in the sample using the Becke line method.
212 8.6 The morphology, relative refractive indices and interference colors can indicate the presence of an
213 explosive ingredient or chemical component. If explosive related components are indicated, additional
214 microscopical analysis is performed to further characterize the components via optical crystallography. If
215 inorganic salts are indicated, additional microscopical analysis can also be performed by microcrystal
216 tests.
217 9. Optical Crystallography by Solvent Recrystallization
218 9.1 Solvent recrystallization allows for a more detailed optical crystallographic characterization of a
219 compound as it recrystallizes from a small drop of suitable solvent.
220 9.2 Water recrystallization for inorganic oxidizers and salts
221 9.2.1 Add 1-3 particles or grains (approximately 1 mg or less) of the specimen to a small drop (5-7 mm in
222 diameter) of distilled or deionized water on a microscope slide. It may be necessary to crush and disperse
223 the sample with the tip of a glass rod. Alternatively, a portion of the specimen can be mixed with water
224 and filtered to remove water insoluble components such as carbon, aluminum, and sulfur; a small drop of
225 the filtered water extract is then placed on a microscope slide.
226 9.2.2 Examine the test drop using the PLM. Crystals typically form at the edge of the drop first.
227 Distorted or subhedral crystals, if formed, can be pushed back into the center of the drop using a glass rod
228 or metal probe to produce characteristic euhedral crystals. Multiple water-soluble components can be
229 present in the sample and, depending on their concentration and solubility, can recrystallize at different
230 times as the drop evaporates.
231 9.2.3 Examine the crystals using plane-polarized and crossed-polarized light. Note their crystal habit,
232 relief, and whether they are isotropic or anisotropic.
233 9.2.4 For anisotropic crystals, determine if extinction is parallel, symmetrical, or oblique.

5
 OSAC 2022-S-0023, Standard Practice for the Forensic
Analysis of Explosives By Polarized Light Microscopy
234 9.2.5 Assess the interference colors to determine if the birefringence is low (less than 0.01), moderate
235 (0.01-0.10), or high (0.10 and above).
236 9.2.6 If observable, use interference figures to determine if the crystal is uniaxial or biaxial and the optic
237 sign. Interference figures can also be used to relate morphology of the crystal to its optical orientation,
238 and, if the crystal is biaxial, estimate 2V.
239 9.2.7 Compare formed crystals to known controls or published references. Table 1 lists optical
240 crystallographic properties for eleven common oxidizers. Document the crystals by description, sketch,
241 or photomicrographs.
242 9.2.8 Dried crystals can be mounted in refractive index oils for refractive index determination, analyzed
243 via microcrystal tests or other instrumental techniques (WK 67862). If the crystals cannot be further
244 analyzed while on the glass slide, use a tungsten needle to transfer the crystals from the glass slide to
245 another substrate, such as a potassium bromide pellet, for additional analysis.
246 9.3 Organic solvent recrystallization for sulfur
247 9.3.1 Add 1-3 particles or grains (approximately 1 mg or less) of sample to a small drop (5-7 mm in
248 diameter) of a suitable solvent such as toluene or methylene chloride. Sulfur, if present, will recrystallize
249 as highly refractive, birefringent dipyramids (diamond-shaped) or form liquid drops that crystallize when
250 touched with a tungsten needle.
251 9.3.2 Document the crystals by description, sketch, or photomicrograph.
252 9.3.3 Compare formed crystals to known controls or published references. Table 1 lists optical
253 crystallographic properties for sulfur.
254 9.3.4 Dried crystals can be further analyzed by other analytical techniques such as Raman or SEM-EDS.
255 If the crystals cannot be further analyzed while on the glass slide, use a tungsten needle to transfer
256 crystals from the glass slide to another substrate, such as carbon tape, for additional analysis.
257 10. Optical Crystallography Using Refractive Index Liquids
258 10.1 Organic and inorganic compounds present in explosives are characterized by their optical
259 crystallographic properties by mounting intact material in a liquid of known refractive index.
260 10.2 Mount 1-3 particles or grains (approximately 1 mg or less) of the specimen, or dried crystals
261 obtained during recrystallization, as described in Section 9, in a refractive index oil. If previous
262 examinations indicate a particular compound, mount in a refractive index oil to confirm one or more
263 known refractive indices of that compound.
264 10.3 Examine using PLM to determine the optical crystallographic properties such as refractive indices,
265 birefringence, extinction type (parallel, symmetrical or oblique), optic sign, and optic axial angle (2V).
266 10.4 Refractive index may be determined using the Becke line method or dispersion staining.
267 10.5 Compare the optical crystallographic properties of the specimen to known reference materials or
268 published references (6, 8, 12, 13, 22). Table 1 lists optical crystallographic properties for sulfur, ascorbic
269 acid, sucrose and twelve common oxidizers. Table 2 lists optical crystallographic properties for four
270 organic high explosives.
271

6
 OSAC 2022-S-0023, Standard Practice for the Forensic
Analysis of Explosives By Polarized Light Microscopy
272 Table 1. Optical Crystallographic Properties of Common Fuels & Oxidizers

Compound Crystal Refractive Habit Comments

System Indices/
Optic
Angle
S (12) orthorhombic 1.958 dipyramids High relief in solvent. Larger crystals
2.038 appear yellowish in color.
2.245
2V = 68
O
(+)
Ascorbic Acid monoclinic 1.474 High order interference colors
(22) 1.595
1.746
2V =
88O(+)
Sucrose (12, monoclinic 1.540
22) 1.567
1.572
2V = 48O(-)
NaClO3 (8) cubic 1.518 squares,
rectangles
Ba(NO3)2 (8) cubic 1.571 cubes
octahedra
Sr(NO3)2 (6) cubic 1.586 octahedra

Pb(NO3)2 (8) cubic 1.781 octahedra High relief in water

cubes
KClO4 (8) orthorhombic 1.4731 rectangular Low order interference colors
1.4737 prisms, rhombs
1.4769
2V =
50O(+)
NH4ClO4 (8) orthorhombic 1.4818 six-sided prisms Low order interference colors. Crystals
1.4833 lying on a {110} face will exhibit an off-
1.4881 centered biaxial interference figure.
2V =
70O(+)
NaClO4 (8) orthorhombic 1.4606 See Sodium chlorate recrystallizes as the
1.4617 NaClO4.2H20 hydrate. Optical data is for the
1.4730 anhydrous.
2V =
69O(+)
NaClO4.2H20 monoclinic diamonds, Low relief in water. Some orientations
(8) rhombs exhibit oblique extinction. Refractive
indices not known.
KClO3 (8) monoclinic 1.415 diamond-shaped Moderate order interference colors,
1.517 tablets, often symmetrical extinction
1.523, truncated

7
 OSAC 2022-S-0023, Standard Practice for the Forensic
Analysis of Explosives By Polarized Light Microscopy
2V = 28O(-)
KNO3 (8) hexagonal & 1.3346 rhombohedra & The hexagonal forms with high order
orthorhombic 1.5056 chevrons early; interference colors appear first at the edge
1.5064 prisms late of the drop. Using a glass rod to move
2V = 7O (-) them to the middle of the drop causes the
stable orthorhombic forms to appear.
NH4NO3 (8, 9) orthorhombic 1.413 prisms, as blades Crystal formation is slow due to the high
1.611 & rods solubility of ammonium nitrate. The
1.637 resulting crystals exhibit high order
2V = 35O (- interference colors. As a fusion
) preparation cools, three to four
polymorphs are observed. MP = 169
NaNO3 (8) hexagonal 1.5874 (ω) rhombohedra High order interference colors and
1.3361 (ε) symmetrical extinction.
273
274 Table 2. Crystallographic and Optical Properties of High Explosives

Compound Crystal System Optical Properties Habit Comments

RDX (13) orthorhombic 1.578 Exhibits anomalous
1.597 interference colors.
1.602 MP = 204-206
2V = 53O(-)
HMX (13) monoclinic 1.589 dipyramids & Typically exhibits high
1.594 pinacoids, rare order interference
1.73 acicular colors.
2V = 20O (+) Oblique extinction

TNT (13) orthorhombic 1.543 MP = 81

1.674
1.717
2V = 60 (-)
PETN (13) tetragonal ω = 1.556 MP = 141.5-142
ε = 1.551
275
276 11. Microcrystal tests
277 11.1 Microcrystal tests are chemical precipitation reactions where the target substance precipitates with
278 the reagent. The crystals formed are examined using a polarized light microscope. Microcrystal tests are
279 used in conjunction with other independent techniques to identify explosive compounds.
280 11.2 Microcrystal tests are used to test for anions and cations of oxidizers and other substances, such as
281 aluminum, that are present in explosives. Many microcrystal tests are available, but those commonly
282 used in explosives analysis are listed in Table 3.
283 11.3 Prior to casework, the reagents used for microcrystal tests shall be tested for reliability using the
284 appropriate chemical standard and negative controls. Once it is determined that the reagents are
285 producing the expected response, the reagents can be used for testing case specimens.

8
 OSAC 2022-S-0023, Standard Practice for the Forensic
Analysis of Explosives By Polarized Light Microscopy
286 11.4 There are four routine methods for conducting microcrystal tests (4). Test drops and reagent drops
287 used for microcrystal tests should be approximately 5-7 mm in diameter or smaller. The reactions
288 normally take place within one minute.
289 11.4.1 Method 1
290 11.4.1.1 Place a drop of deionized water on a clean microscope slide and add one or more particles of the
291 sample with stirring, crushing and dispersing as needed. Keep this test drop small and from spreading on
292 the slide.
293 11.4.1.2 A drop of reagent is placed near, but not touching, the test drop. Depending on the microcrystal
294 test being conducted some reagent solutions are pre-made and a drop added to the slide. Other reagent
295 drops are created on the slide by placing a drop of water near the test drop and adding 1-5 particles of the
296 reagent to the water droplet.
297 11.4.1.3 Place the slide on the microscope stage.
298 11.4.1.4 Using a clean glass rod, draw one drop into the other drop and immediately examine the
299 preparation microscopically using a 10X objective. Higher magnification may be used but care must be
300 taken so the objective does not contact the test drop.
301 11.4.1.7 Observe the entire preparation including the mixing zone and within each drop. Note the
302 presence of characteristic microcrystals or the lack of a reaction.
303 11.4.1.8 The formation of microcrystals corresponding to those obtained with the chemical standard is
304 indicative for that anion or cation.
305
306 11.4.2 Method 2
307 11.4.2.1 Follow 11.4.1 except add the reagent drop into the middle of the test drop.
308 11.4.2.2 Observe the preparation microscopically as described in Method 1. Note the presence of
309 characteristic microcrystals or the lack of a reaction.
310 11.4.2.3 The formation of microcrystals corresponding to those obtained with the chemical standard is
311 indicative for that anion or cation.
312
313 11.4.3 Method 3
314 11.4.3.1 Follow 11.4.1 except add reagent particles into the middle of the test drop.
315 11.4.3.2 Observe the preparation microscopically as described in Method 1. Note the presence of
316 characteristic microcrystals or the lack of a reaction.
317 11.4.3.3The formation of microcrystals corresponding to those obtained with the chemical standard is
318 indicative for that anion or cation.
319
320 11.4.4 Method 4
321 11.4.4.1. Place a small drop of deionized water on a clean microscope slide and add one or more particles
322 of the sample with stirring, crushing, and dispersing as needed. Keep this test drop small and from
323 spreading on slide.
324 11.4.4.2 Place a glass ring with an internal diameter of about 10-12 mm around the test drop so the drop is
325 in the center of the ring.
326 11.4.4.3 Place a small drop of prepared reagent on a coverslip.
327 11.4.4.5 Place the slide on a microscope stage making sure there is sufficient working distance between
328 the top of the glass ring and the objective.

9
 OSAC 2022-S-0023, Standard Practice for the Forensic
Analysis of Explosives By Polarized Light Microscopy
329 11.4.4.6 Add a small drop of dilute (approximately 10-30%) sodium hydroxide to the test drop inside the
330 center of the ring.
331 11.4.4.7 Immediately place the coverslip onto the glass ring so the reagent drop is suspended in the center
332 of the ring above the sample.
333 11.4.4.8 Observe the reagent drop for the presence of microcrystals or the lack of reaction.
334 11.4.4.9 The formation of microcrystals corresponding to those obtained with the chemical standard is
335 indicative for that anion or cation.
336
337 11.5 Table 3 lists twelve tests for anions and cations common to low explosives along with the reagent,
338 literature reference, and method as described in 11.4. Microcrystal tests for additional ions can be found
339 in Chamot & Mason (4), Benedetti-Pichler (21), Wills (19), and Hollifield (20).
340
341 Table 3. Microcrystal Tests for Ions

Cation/anion tested Microcrystal test & reference Method

Potassium Chloroplatinic acid (4) 1, if positive, repeat using
Method 4, if potassium present
and ammonium ion absent
method 4 test will be negative
Ammonium Chloroplatinic acid (4) 4
Sodium zinc/uranyl acetate (4) 4
Aluminum ammonium molybdate (4) 2
Magnesium Squaric acid (19, 20) 2
Barium Squaric acid (19, 20) 2
Strontium Squaric acid (19, 20) 2
Lead Potassium iodide (4) 1
Nitrate Nitron sulfate (1, 4) 1
Perchlorate Strychnine sulfate (4) 1, 3
Perchlorate Methylene Blue (7) 1
Chlorate Methylene Blue (7) 1
342
343 12. Fusion Methods
344 12.1 Fusion methods (10) are techniques that allow for the characterization of crystals by determining the
345 melting point and observing optical crystallographic properties as the sample recrystallizes from the melt.
346 This method is most useful for organic high explosives (11, 14, 15, 16) but can also be applied to some
347 inorganic explosives (9).
348 12.2 The solid sample is placed on a microscope slide with a cover slip, or on a cover slip (according to
349 manufacturer’s instructions), placed in the hot stage, and brought into the field of view. The temperature
350 in the hot stage is raised as the sample is observed using the polarized light microscope. The melting
351 point is documented and the temperature is lowered slowly while observing the recrystallization of the
352 sample from the melt.
353 12.3 Note the optical crystallographic properties such as morphology, crystal front shape, polymorphism,
354 interference colors, anomalous interference colors, extinction characteristics, interference figure, and
355 optic sign for each solid state as appropriate.

10
 OSAC 2022-S-0023, Standard Practice for the Forensic
Analysis of Explosives By Polarized Light Microscopy
356 12.4 Most inorganic oxidizers found in low explosives have melting points above 300°C. These higher
357 melting oxidizers are heated on a microscope slide using an alcohol lamp (9). An accurate melting point
358 is not determined, but the sample is observed through the microscope as the slide cools to obtain various
359 optical crystallographic properties, as described in 12.3.
360 12.5 Compare the optical crystallographic properties of the specimen to published reference
361 descriptions (9-11, 14-16) or a known reference material.
362 12.6 Mixed fusions
363 12.6.1 If an explosive compound is indicated using optical crystallography, microcrystal tests, or other
364 techniques, a mixed fusion using a known reference material of the suspected compound can confirm the
365 identity of the unknown (9, 10).
366
367 12.6.2 A small amount of material from the specimen is placed near the edge of a coverslip, melted over
368 an alcohol lamp, and allowed to flow under part of the coverslip. Limit the amount of specimen so that
369 the entire area under the coverslip is not filled with the specimen.
370 12.6.3 Once the unknown specimen has cooled and recrystallized, the known reference material is placed
371 near the opposite edge of the coverslip and melted over an alcohol lamp such that the melted standard
372 flows under the coverslip and contacts the previously cooled unknown specimen. The portion of the
373 unknown specimen that contacts the reference material will also melt during this process.
374 12.6.4 Using the PLM, view the zone of mixing as the preparation cools and recrystallizes. If the
375 unknown compound recrystallizes into the reference material without differences in velocity, appearance,
376 or the presence of a eutectic, this is strong evidence that the two compounds are the same.
377 12.6.5 Once cooled, the mixed fusion preparation can be inserted into the hot stage at a temperature
378 approximately 10 degrees lower than the melting point of the reference material. While observing the
379 zone of mixing, slowly raise the temperature (1-3 degrees/minute) of the hot stage to the melting point of
380 the reference material If the two compounds are the same, the melting point will be the same across
381 the zone of mixing. Note: this can only be performed if the sample melts below the upper temperature
382 range of the hot stage.
383 13. Documentation
384 13.1 All details necessary to support the interpretation made from the examinations are documented.
385 13.2 Examination notes are reviewed to ensure they reflect the particular characteristics used in the
386 microscopical examination including measured, observed, or calculated values, descriptions, sketches, or
387 photomicrographs.
388 13.3 Examination documentation is reviewed to ensure they contain sufficient detail to support the
389 conclusions such that another qualified examiner could independently reach the same conclusion based on
390 the notes or documentation.
391 14. References
392 1. Lawrence, G., and Fink, M., “The Evaluation of Nitron Sulfate as a Microchemical Test for Some
393 Common Oxidizers,” The Microscope, Vol 55, No. 2, 2007, pp 55-58.

11
 OSAC 2022-S-0023, Standard Practice for the Forensic
Analysis of Explosives By Polarized Light Microscopy
394 2. Parker, R. G., Stephenson, M. O., McOwen, J. M., and Cherolis J. A., “Analysis of Explosives and
395 Explosive Residues. Part 1: Chemical Tests, ” Journal of Forensic Science, Vol 20, No. 1, January 1975.
396 3. Chamot E. M., and Mason C. W., Handbook of Chemical Microscopy, Volume 1 John Wiley & Sons,
397 Inc., New York, NY, 1958, p. 51.
398 4. Chamot, E., and Mason, C., Handbook of Chemical Microscopy, Volume II, John Wiley & Sons, Inc.,
399 New York, NY, 1940.
400 5.Hopen, T. J., “Tricks of the Trade, Extraction and Identification of Sulfur,” The Microscope, Vol 48,
401 No. 1, 2000, p. 51.
402 6. Winchell A. N., The Microscopic Characters of Artificial Inorganic Solid Substances or Artificial
403 Minerals, John Wiley & Sons, Inc., New York, NY, 1931.
404 7. Hopen T. J., and Crippin J. B., “Methylene Blue Microchemical for the Detection and Identification of
405 Chlorates and Perchlorates,” The Microscope, Vol 49, 2001.
406 8. Hopen T.J., and Kilbourn J. H., “Characterization and Identification of Water Soluble Explosives,” The
407 Microscope, Vol 33, No. 1, 1985.
408 9. Kilbourn J. H., and McCrone W. C., “Fusion Methods Identification of Inorganic Explosives,” The
409 Microscope, Vol 33, No. 2, 1985.
410 10. McCrone W. C., Fusion Methods in Chemical Microscopy: A Textbook and Laboratory Manual,
411 Interscience Publishers, Inc., New York, NY, 1957.
412 11. McCrone W. C., “The Identification of High Explosives by Microscopic Fusion Methods,”
413 Microchemical Journal, Vol 3, 1959, pp 479-490.
414 12. McCrone W. C., and Delly J. G., The Particle Atlas, Edition 2, Volumes I, II & IV, Ann Arbor Science
415 Publishers, Inc., Ann Arbor, MI, 1973.
416 13. McCrone W. C., Delly J. G., and Palenik S.J., The Particle Atlas, Edition 2, Volume V, Ann Arbor
417 Science Publishers, Inc., Ann Arbor, MI, 1979.
418 14. McCrone W. C., Andreen J. H., and Tsang S. M. “Identification of Organic High Explosives,” The
419 Microscope, Vol 41, 1993, pp 161-182.
420 15. McCrone W. C., Andreen J. H., and Tsang S. M. “Identification of Organic High Explosives, II,” The
421 Microscope, Vol 42, 1994, pp 61-73.
422 16. McCrone W. C., Andreen J. H., and Tsang S. M. “Identification of Organic High Explosives III,” The
423 Microscope, Vol 42, 1999, pp 183-200.
424 17. McCrone, W.C., “Solubility, Recrystallization and Microchemical Tests on Nanogram Single
425 Particles,” The Microscope, V34 N2, 1986, pp 107-118.
426 18. Delly J. G., Essentials of Polarized Light Microscopy and Ancillary Techniques, Hooke College of
427 Applied Sciences, Westmont, IL, 2017.
428 19. Wills WF, Jr., “Squaric Acid Revisited,” The Microscope, Vol 38, 1990.

12
 OSAC 2022-S-0023, Standard Practice for the Forensic
Analysis of Explosives By Polarized Light Microscopy
429 20. Hollifield, J.M., “Characterization of Squaric Acid Precipitates,” The Microscope, V51 N2, 2003, pp
430 81-103.
431 21. Benedetti-Pichler, A.A., “Identification of Materials via Physical Properties, Chemical Tests, and
432 Microscopy”, Academic Press Inc., New York, NY, 1964.
433 22. Winchell A. N., The Optical Properties of Organic Compounds, McCrone Research Institute,
434 Chicago, IL, 1987 (reprint of 1942 edition, Academic Press, Inc)
435 23. New York Microscopical Society, Glossary of Microscopical Terms and Definitions, Second Edition,
436 McCrone Research Institute, Chicago, IL, 1989.

13