Manual of basic techniques for a health laboratory - libdoc.who.int

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352 Manual of basic techniques for a health laboratoryAlkaline haematin D reagent (No. 8)Sodium hydroxide (NaOH)4 gTriton X-100 (or equivalent)25gDistilled water1000mlDissolve the sodium hydroxide in the distilled water in a clean conical flask. Stirusing a glass rod until the crystals have completely dissolved. Add the Triton X-100(or equivalent) and mix well. Filter the solution into a clean glass-stoppered reagentbottle, using Whatman No. 1 (or equivalent) filter-paper. Label the bottle“ALKALINE HAEMATIN D REAGENT” and write the date. Store at room temperature(20–25°C).Check the quality of the solution (see below).Alkaline haematin D (AHD) reagent will keep for several months at 20–25°C. If aprecipitate forms during storage, the reagent should be filtered before use.Note: Use filtered rainwater if distilled water is not available.Quality control of alkaline haematin D reagentAn alkaline haematin D standard solution (AHD standard) supplied by the centrallaboratory is used to test the quality of new batches of AHD reagent in peripherallevellaboratories.1. Fill a clean cuvette with distilled water. Place the cuvette in the cuvette chamberand adjust the haemoglobinometer or colorimeter to read zero at 540nmwavelength.2. Replace the distilled water with AHD reagent. The haemoglobinometer or colorimetershould read zero.3. Pipette 20ml of AHD standard into a test-tube containing 3ml of the freshlyprepared AHD reagent (1:150 dilution).4. Measure the haemoglobin concentration of the AHD standard (see section 9.3.2).5. Repeat the procedure using the previous batch of AHD reagent. Compare theresults.6. If the haemoglobin values differ by more than 5g/l, discard the freshly preparedAHD reagent and prepare a new batch, paying attention to accurate measurementof the constituents and the cleanliness of the glassware.The AHD standard stock solution will keep for 8 months at 4–8°C.Amies transport medium (No. 9)Charcoal, pharmaceutical grade 10.0gSodium chloride (NaCl) 3.0gDisodium hydrogen phosphate (Na 2HPO 4·2H 2O) 1.15gPotassium dihydrogen phosphate (KH 2PO 4) 0.20gSodium thioglycollate 0.10gCalcium chloride (CaCl 2), anhydrous 0.10gMagnesium chloride (MgCl 2) 0.10gAgar 4.00gDistilled water1000mlSuspend the mixture of salts in the distilled water. Add the agar and heat until theagar has completely dissolved. Add the charcoal. Dispense into small tubes or bottleswhile stirring to keep the charcoal evenly suspended.
Annex. Reagents and their preparation 353Sterilize by autoclaving at 120°C for 15min. Cool immediately in cold water tokeep the charcoal evenly suspended. Label the tubes or bottles “AMIES TRANS-PORT MEDIUM” and write the date.Amies transport medium is also available commercially.Benedict solution (No. 10)Copper sulfate (CuSO 4·5H 2O) 17.3gTrisodium citrate (Na 3C 6H 5O 7·2H 2O)173gSodium carbonate (Na 2CO 3), anhydrous100gDistilled waterq.s. 1000mlDissolve the copper sulfate crystals by heating in 100ml of distilled water.Dissolve the trisodium citrate and the sodium carbonate in about 800ml ofdistilled water. Add the copper sulfate solution slowly to the sodium carbonate/trisodium citrate solution, stirring constantly. Make up the volume to 1000ml withdistilled water. Transfer the solution to a glass-stoppered bottle. Label the bottle“BENEDICT SOLUTION” and write the date.Blank reagent (No. 11)Trichloroacetic acid (CCl 3COOH), 50g/l (5%) solution(5g in 100ml of distilled water; see No. 62) 50mlDistilled waterq.s. 100mlMix. Transfer the solution to a glass-stoppered bottle. Label the bottle “BLANKREAGENT” and write the date. Store at room temperature (20–25°C).The reagent will keep for several months at 20–25°C.Warning: Trichloroacetic acid is highly corrosive.Boric acid, saturated solution (No. 12)Boric acid 4.8gDistilled waterq.s. 1000mlStore in a glass-stoppered bottle. Label the bottle “BORIC ACID SATURATEDSOLUTION” and write the date.Brilliant cresyl blue (No. 13)Brilliant cresyl blue 1.0gTrisodium citrate (Na 3C 6H 5O 7·2H 2O) 0.4gSodium chloride (NaCl), 8.5g/l (0.85%) solution(No. 53)100mlDissolve the dye and the trisodium citrate together in the sodium chloride solution.Filter the solution obtained into a staining bottle. Label the bottle “BRILLIANTCRESYL BLUE” and write the date.Buffered glycerol saline (No. 14)Sodium chloride (NaCl) 4.2gDipotassium hydrogen phosphate (K 2HPO 4),anhydrous 3.1gPotassium dihydrogen phosphate (KH 2PO 4),anhydrous 1.0gPhenol red 0.003g
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M A N U A LO F B A S I CTECHNIQUESF
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ContentsiManualofbasic techniques f
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ContentsiiiContentsPrefacex1. Intro
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viManual of basic techniques for a
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viiiManual of basic techniques for
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xManual of basic techniques for a h
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Table 3.2 Dispatch of specimens 92
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7. Examination of urine 231Part III
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7. Examination of urine 2337. Exami
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7. Examination of urine 2357.2.3 Me
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7. Examination of urine 237Table 7.
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7. Examination of urine 239The anal
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7. Examination of urine 241MethodCo
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7. Examination of urine 243Table 7.
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7. Examination of urine 245Fig. 7.2
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7. Examination of urine 2497.2.8 De
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7. Examination of urine 251Reuse of
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8. Examination of cerebrospinal flu
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9. Haematology 2659. HaematologyHae
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9. Haematology 2679.2 Collection of
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9. Haematology 269Fig. 9.13 Feeling
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9. Haematology 2719.3 Estimation of
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9. Haematology 273Table 9.2 Sample
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9. Haematology 275Fig. 9.25 Checkin
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9. Haematology 277Calibration of th
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9. Haematology 279Errors in haemogl
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9. Haematology 281Fig. 9.28 Micro s
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9. Haematology 283Fig. 9.34 Centrif
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9. Haematology 285concentration is
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9. Haematology 287Measurement techn
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9. Haematology 289●●●Graduate
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9. Haematology 2911 litre, so multi
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9. Haematology 293Fig. 9.45 Materia
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9. Haematology 295Very high ESR val
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9. Haematology 2979.9 Observation o
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9. Haematology 2999.10 Preparation
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Page 331 and 332: 9. Haematology 319Heinz bodiesHeinz
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Page 341 and 342: 11. Immunological and serological t
Page 363: Annex. Reagents and their preparati
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Page 381 and 382: IndexNote: Page numbers in bold ref
Page 383 and 384: Index 371Centrifuges 32, 69-72, 69-
Page 385 and 386: Index 373Endolimax nanus 114, 114,
Page 387 and 388: Index 375Hyaline casts, urine 243,
Page 389 and 390: Index 377Microhaematocrit equipment
Page 391 and 392: Index 379Protozoablood 173-194, 178
Page 393 and 394: Index 381Smears (continued)preparat
Page 395 and 396: Index 383Thiomersal-iodine-formalde
Page 397 and 398: Selected WHO publications of relate
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