Page:NIOSH Manual of Analytical Methods - 7400.pdf/5

 ASBESTOS and OTHER FIBERS by PCM: METHOD 7400, Issue 2, dated 15 August 1994 - Page 5 of 15

acetone into the vaporization chamber with a slow, steady pressure on the plunger button while holding pipet firmly in place. After waiting 3 to 5 s for the filter to clear, remove pipet and slide from their ports. CAUTION:	Although the volume of acetone used is small, use safety precautions. Work in a well-ventilated area (e.g., laboratory fume hood). Take care not to ignite the acetone. Continuous use of this device in an unventilated space may produce explosive acetone vapor concentrations. c.	 Using the 5-µL micropipet, immediately place 3.0 to 3.5 µL triacetin on the wedge. Gently lower a clean cover slip onto the wedge at a slight angle to reduce bubble formation. Avoid excess pressure and movement of the cover glass. NOTE:	If too many bubbles form or the amount of triacetin is insufficient, the cover slip may become detached within a few hours. If excessive triacetin remains at the edge of the filter under the cover slip, fiber migration may occur. d.	 Mark the outline of the filter segment with a glass marking pen to aid in microscopic evaluation. e.	 Glue the edges of the cover slip to the slide using lacquer or nail polish [12]. Counting may proceed immediately after clearing and mounting are completed. NOTE:	If clearing is slow, warm the slide on a hotplate (surface temperature 50 °C) for up to 15 min to hasten clearing. Heat carefully to prevent gas bubble formation. CALIBRATION AND QUALITY CONTROL: 10.	Microscope adjustments. Follow the manufacturer’s instructions. At least once daily use the telescope ocular (or Bertrand lens, for some microscopes) supplied by the manufacturer to ensure that the phase rings (annular diaphragm and phase-shifting elements) are concentric. With each microscope, keep a logbook in which to record the dates of microscope cleanings and major servicing. a.	 Each time a sample is examined, do the following: (1)	Adjust the light source for even illumination across the field of view at the condenser iris. Use Kohler illumination, if available. With some microscopes, the illumination may have to be set up with bright field optics rather than phase contract optics. (2)	Focus on the particulate material to be examined. (3)	Make sure that the field iris is in focus, centered on the sample, and open only enough to fully illuminate the field of view. b.	 Check the phase-shift detection limit of the microscope periodically for each analyst/microscope combination: (1)	Center the HSE/NPL phase-contrast test slide under the phase objective. (2)	Bring the blocks of grooved lines into focus in the graticule area. NOTE:	The slide contains seven blocks of grooves (ca. 20 grooves per block) in descending order of visibility. For asbestos counting, the microscope optics must completely resolve the grooved lines in block 3 although they may appear somewhat faint, and the grooved lines in blocks 6 and 7 must be invisible when centered in the graticule area. Blocks 4 and 5 must be at least partially visible but may vary slightly in visibility between microscopes. A microscope which fails to meet these requirements has resolution either too low or too high for fiber counting. (3)	If image quality deteriorates, clean the microscope optics. If the problem persists, consult the microscope manufacturer. 11.	Document the laboratory’s precision for each counter for replicate fiber counts. a.	 Maintain as part of the laboratory quality assurance program a set of reference slides to be used on a daily basis [13]. These slides should consist of filter preparations including a range of loadings and background dust levels from a variety of sources including both field and reference samples (e.g., PAT, AAR, commercial samples). The Quality Assurance Officer should maintain custody of the reference slides and should supply each counter with a minimum of one reference NIOSH Manual of Analytical Methods (NMAM), Fourth Edition