Andrology Rotation MC

What do you do if CEROS QC is not in range?

What are three things you do if CEROS QC repeat reads are unacceptable?

Who are two people that need to be contacted if the analyzer is having issues?

Notify other shifts/techs and if not able to be remedied by troubleshooting, notify repair or maintenance

When is a photo ID necessary for patients?

When a sample is used for IUI or other procedure involving use of a human product, a government issued ID is required to identify and confirm the

What specimen must be kept refrigerated?

What specimen must be kept at room temperature?

What is considered abnormal viscosity?

a sample droplet producing a thread length greater than 2 cm

What are gel particles?

Large clumps of unliquified sample that sediment at the bottom of a sample and contain no sperm cells

When are dilutions made of the sample?

For a manual count, a dilution is performed when there is a high concentration and counting is difficult; dilution is performed to simplify counting

For CEROS, too high of a concentration would also make it difficult for the analyzer to identify individual cells

Why is mechanical liquification performed?

When a sample does not liquify after 30 minutes and with aspiration with a 20g needle

• continue stair stepping aspiration with smaller needle gauges

• add media and saline to thin out the sample

What are some other factors that could be notated as abnormal?

• color

• agglutination/clumping

• viscosity

What is sperm-to-sperm agglutination?

Sperm agglutinating with other sperm cells, often motile

What are sperm cell clumps?

Sperm cells are clumping with other cells or other debris

When would you spin post-vasectomy samples?

If 10 or more sperm cells are seen in 5 fields on a neat sample, how is this reported?

Concentrating of the sample by centrifugation is not required and the concentration calculated can be reported.

• also known as 5 n’ out

What is the minimum amount of time a fresh-collected sample must sit prior to analysis?

15 minutes

How do you measure/count when there is little to no sample in the collection container?

Add media

How do you correct values for samples arriving in media?

Media is aliquoted at 4 mL per sample. The difference in the volume is the sample volume.

If the first two manual counts do not match; what is done next?

Perform a third count and whichever of the first two counts is closest or within range of the third count is used in the average.

Two of the three manual counts do not match; what three things could be wrong?

• Sample clumping/viscosity

• Slide is not functioning properly

• Natural variability

Two of the three manual counts on extremely low counts do not match after troubleshooting; how would you report?

What are the four reasons a manual count would be performed over a CEROS analysis?

1. motility <5%

2. count <1

Why is a visual estimate performed when reading samples on CEROS?

How do you adjust CEROS gates and why?

How many fields and cells are read on CEROS depending on the sample?

• samples with <400 cells need 5 fields counted at least

• samples with high counts need a minimum of 400 cells in no less than 3 fields

When is a third reading performed with CEROS?

What are the steps and timing for staining a slide?

Fixative - 15 secs

Stain A - 10 secs

Stain B - 5 secs

Distilled water - 15 secs

How is quality of stain checked?

Visual examination

What does a slide with poor stain quality look like?

Too dark or too light of a stain where visibility of the sperm cells are not clear due to lightness of staining or debris from over staining

When are slides done on diluted samples and why?

Which patients require a Consent to use form on file and why?

Donor patients require this to allow their samples to be used for IUI or other procedures

Where are two places to check for a Consent to use file?

Why record CMV status and why must a provider sign off on it?

CMV can cause fetal abnormalities in pregnancy and is required that the patient if CMV = is aware and consents to the use of the sample. Physicians need to sign to ensure that the patient has been informed.

How do you troubleshoot a “fluffy pellet”?

Rewash the cells and liquify them better before trying to recentrifuge the sample to ensure no streaks in the supernatant

If neat total count is >100 M/mL what needs to be done for the spun sample?

A dilution

If the sample has antibodies to sperm, what needs to be done?

What is the latest date a cancer patient can freeze prior to treatment

Anytime before treatment is valid so long as the physician allows it

Why is a pre-count done prior to freezing?

To determine the amount of cells to be frozen is in the solution as well as determine a baseline for the post-freeze analysis

Why is a count done after freezing?

To determine the survivability of the sample and how many are still viable

What is the minimum time for sample and reagent to equilibrate prior to freezing?

How do cryoprotectants minimize intracellular ice crystals and cell damage?

They displace water within the cell which lowers the risk of the cell bursting due to ice crystal formation when water freezes

What evidence needs to be provided to show that samples are frozen properly?

Time that the sample is out into each freezer as well as a post-freeze analysis to see the survivability of the cells

What is the difference in communicable disease testing for SIP and donor samples?

Regulations for testing are not applied to sexual intimate partners (SIP) and testing performed for donors includes:

• HIV 1/2

• HBV - HBsAG and anti-HBc

• HCV

• Syphilis

• anti-HTLV 1/2

• anti-CMV

• Chlamydia trachomatis

• Neisseria gonorrhea

For incoming donor samples, what information is captured for FDA purposes?

• sample is maintained at acceptable temperature range

• no compromise of container or device

• donor samples have been tested according to FDA requirements

• donor eligibility is documented

• donor summary of records is available

• donor CMV status is documented

Which donor samples are stored in a separate LN₂ tank and why?

CMV positive samples are put into separate tanks because of the potential that there will be cross contamination of samples

How is chain-of-custody handled with frozen samples entering or exiting the lab?

Must be maintained for all HCT/P samples including autologous/SIP and must include:

• date

• time

• individual handling the sample

• sample destination/use