Appendix 2: Filling The Sqa-V Capillary With A Low Volume Sample - MES SQA-V Gold User Manual

User Guide Version 2.60 I-button WHO 5th

APPENDIX 2: Filling the SQA-V Capillary with a Low Volume Sample

Sample size, collection container and preparation:
1. A sample as small as 20 micro liters can be tested for motility parameters by filling ONLY the thin
section of the testing capillary (Figure 1).
2. The semen sample must be completely liquefied and well mixed prior to
aspiration. Gently rotate the container to fully mix the liquefied specimen.
WARNING: Do not shake nor use a pipette to aspirate and dispense
specimen in order to mix, otherwise air bubbles will form.
3. Carefully check that the liquefied, fully mixed specimen is free of air
bubbles (or that there is an adequate amount of sample below the air
bubbles) before immersing the capillary into the specimen, thus ensuring that
no air bubbles will be aspirated into the capillary.
4. It is recommended that the sample be withdrawn from a standard
"tissue culture dish" (3 cm in diameter/1 cm deep) to allow for better
visual control when filling the capillary.
Figure 2
Filling the capillary:
1. Push the syringe piston in fully. Place only the thin part of the capillary
into the bottom of the sample (Figure 1).
2. Pull the piston back slowly without withdrawing the capillary from the
sample. Fill only the (thin) capillary chamber with 20 micro liters of
semen (Figure 1). The exact quantity aspirated can be determined by the
gradations on the 1 ml syringe. Aspirate the sample until it just appears in
the cuvette part while keeping the tip of the capillary well below the sample
level and well below the level of any bubbles covering the liquid. Withdraw
the capillary tip from the semen sample and visually inspect the capillary to
ensure that the sample has completely filled the thin section (no meniscus).
3. Quickly (to avoid wicking) and thoroughly wipe the outer surface of the
capillary - both top and bottom with a delicate wipe (Kimwipes, etc.). It is
important to remove all semen from the exterior of the capillary in order to
prevent the SQA-V optical chamber from becoming clogged. Visually confirm
that the thin chamber of the capillary is still full of semen after completing
the cleaning process. If some of the sample has been depleted push-in the
piston slightly until the first drop appears on the capillary tip and then fill the
capillary again from the sample container.
4. The separating valve must now be removed. Detach the entire syringe from the hub (Figure 2) and
use the syringe tip to firmly push-out the separating valve from the underside of the capillary
(Figure 3). Completely detach the separating valve (Figure 4). The capillary is now ready to be
inserted into the SQA-V.
5. PLEASE NOTE:
capillary.
Revision 21_Sep_2015
Test Low Volume samples as soon as the sample is aspirated into the
33
Figure 1
Figure 3
Figure 4