A Methods Manual for the Collection, Preparation and Analysis of ...

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1: Micro-pipette 1 ml with disposable tips 2: Glass and plastic Pasteur-pipettes (2-3 ml). 3: Forceps.4: Microscope slides. 5: Coverslips. 6: Diatom mountant. 7: 10 ml plastic gradated centrifuge tube.8: 15 ml glass test tube. 9: 4 ml glass sample storage bottles with rubber seal inside cap. 10: Waterprooffine marking pen. 11: Heat-resistant glass beaker ~ 100 ml. 12: Watch glass.27
Section SummaryPreparation:1. Pre-preparation examination for live cells.2. Cleaning of cells:a. In laboratory equipped with a fume cabinet: KMnO 4 + hot HClmethod, hot H 2 SO 4 + HNO 3 (2:1) method, hot H 2 O 2 method.b. In WELL VENTILATED laboratory without fume cabinet: ColdH 2 O 2 .c. Rinsing: Centrifuge available: centrifuge with distilled water untilsample is circumneutral (4-5 runs for 10 min. at 2500 rpm)d. No centrifuge available: decant supernatant using an aspirator.Resuspend sample using distilled water and allow to settle for 8 hours(repeat 4-5 times)3. Slide preparation:a. Concentrated diatom solution diluted with distilled water until onlyslightly cloudy,b. Add 1-2 drops of 10% NH 4 Cl to dilute solution to prevent clumpingof cells,c. 1.5 – 2 ml of dilute solution is placed on cover slip, depending on sizeof cover slip,d. Sample allowed to air-dry ( Takes approximately 24 hrs),e. Cover slip heated to drive off excess moisture and sublimate NH 4 Cl,f. Sample mounted with high-resolution mountant.Archiving:1. Cleaned samples should be stored in ethanol, at a concentration high enoughto prevent the growth of bacteria and fungi and to prevent the dissolution ofsilica.2. Slides should be stored flat until mountant is dry.3. All relevant information on the location, date of collection, substratum andcollector should be stored both with the sample and the slide, not simply areference number.28
Page 1 and 2: A Methods Manual for theCollection,
Page 3 and 4: This report is part of a set on Dia
Page 5 and 6: FOREWORDThis method manual is inten
Page 7 and 8: TABLE OF CONTENTSNOTE TO THE USER .
Page 9 and 10: GLOSSARYAerophilic - Depending on f
Page 11 and 12: Phytoplankton - The phytoplankton c
Page 13 and 14: photomicrographs earned high praise
Page 15 and 16: 2. How do you recognise diatoms in
Page 17 and 18: 4. Diatoms - Colony formation and a
Page 19 and 20: 5.1 Pennate and Centric diatomsPenn
Page 22 and 23: SECTION 2: FIELD PROCEDURESTo facil
Page 24 and 25: depending on the period of exposure
Page 26 and 27: 2.3.2 Examination of fresh material
Page 28 and 29: 3. Toolkit for Fieldwork (field app
Page 30 and 31: 5. Site selection for water quality
Page 32 and 33: 6. Sampling locality details and fi
Page 34 and 35: 8. Choice of substrata (detail)Samp
Page 36 and 37: 10 Preservation of diatom material
Page 40 and 41: 1.2 Pre-preparation examination of
Page 42 and 43: 1.3.2.3 Shake the sample well and p
Page 44 and 45: one is available the peroxide can b
Page 46 and 47: 2 Preparation of diatom slidesHealt
Page 48 and 49: 2.1.9 After the diatom-coated cover
Page 50 and 51: 4. ArchivingIt is important to reta
Page 52 and 53: affected at counts of 300 and above
Page 54 and 55: 6. Counting records (electronic and
Page 56 and 57: The main source of variability is,
Page 58 and 59: 11. Key reference worksThere are se
Page 60: HARDING WR, ARCHIBALD CGM, TAYLOR J

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