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Appendix KOVA Counting Procedure

This document provides instructions for using a KOVA-Chamber to manually examine urine samples under a microscope. It describes: 1) The materials needed, including an Eppendorf pipette, KOVA Chamber, and uncentrifuged urine or control sample. 2) How to prepare the chamber by pipetting 10μl of sample and letting it stand for one minute before counting. 3) How to count various particles like red blood cells, white blood cells, and epithelial cells at different magnifications, and how to calculate the results.

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206 views5 pages

Appendix KOVA Counting Procedure

This document provides instructions for using a KOVA-Chamber to manually examine urine samples under a microscope. It describes: 1) The materials needed, including an Eppendorf pipette, KOVA Chamber, and uncentrifuged urine or control sample. 2) How to prepare the chamber by pipetting 10μl of sample and letting it stand for one minute before counting. 3) How to count various particles like red blood cells, white blood cells, and epithelial cells at different magnifications, and how to calculate the results.

Uploaded by

DanielIvanGuerrero
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Procedure KOVA-Chamber

Material

 Eppendorf pipette and pipette tips


 Kova Chamber (do not touch the surface)
 Uncentrifuged urine or control

Chamber preaparation

 Fill the KOVA-Chamber by pipetting 10μl of the sample, with an accurate Eppendorf
pipette, into the small opening. Avoid air bubbles, discard incorrect filled chambers.
 Let the chamber stand for one minute before counting.
 Get an overview of the urine content by scanning the whole chamber (9x9 grids) at low
magnification (100x).
 Count the particles in the grids and the particles touching the left and the lower border of
the grid. Do not count the particles touching the upper and the right border of the grid.
Do not count particles under the wide outer grid-lines if they do not touch the inner
boarder. (Count, do not count)
 Two chambers are counted by two investigators (using a discussion microscope). Build
the mean of each chamber and afterwards build the average over these both single
chambers  calculate the final result as described below.
 Write the number of counted grids, the interim means and the final results in the form:
“Manual microscopy with KOVA-chamber”.

Counting of particles

 Count HYA, SEC and PAT at low magnification (100x) inside the complete grid (81
small grids 0.9 μl).  Multiply by 1.1 = cells/μl

 Count RBC, WBC and NEC at high magnification (400x).


(<5 particles/small grid): Count 36 small grids (4 quadrants)
Multiply by 2.5 = cells/μl
(>=5 particles/small grid): Count 10 small grids (in different quadrants of the
counting area). Multiply by 9 = cells/μl
 For rare particles: Count all 81 grids. Multiply by 1.1 = cells/μl)

Roche Diagnostics International Ltd.


RPD Clinical Operations 19-Sept-2013
 Do not count cells with unusual distribution e.g. clumps. As an exception WBC-clumps
are counted if they are identified reliably (corresponding to the cobas u 701).
 Fill the results in the form: “Manual microscopy with KOVA-chamber”.
 For CRY, YEA, MUC and SPRM rate “pos” if you find some. Write what kind of crystals
you see. Especially Urates are important. For samples providing a concentration of
crystals <=10 p/μL the obtained result has to be recorded quantitatively. Higher
concentrations have to be stated positive only.
 BAC are estimated by using the below shown example pictures. Use: neg, +, ++, +++

 Please note that single mucus strings are recorded as negative. Please find below examples
about negative and positive results.

 Please note that yeast is mostly seen in chains and not that often found as single cells but
the rating is only positive or negative. There will be no counting of the yeast cells
performed. Please find below examples about negative and positive results.

Roche Diagnostics International Ltd.


RPD Clinical Operations 19-Sept-2013
Bacteria

1+

2+

3+
Roche Diagnostics International Ltd.
RPD Clinical Operations 19-Sept-2013
Mucus

Single mucus
string; rated as
negative

Many mucus
strings; rated as
positive

Roche Diagnostics International Ltd.


RPD Clinical Operations 19-Sept-2013
Yeast
Single yeast cells in
one image will be
rated as negative, but
single cells found in
several images is to be
rated positive.

Many single yeasts and chains;


rated as positive

Roche Diagnostics International Ltd.


RPD Clinical Operations 19-Sept-2013

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