Here is a question that has came up:

What is the difference between Chymotrypsin and Hyaluronidase?

Answer:

In the article: Stefan Stephanus du Plessis, Sheila Gokul, Ashok Agarwal. Frontiers in Bioscience, Elite, 5, 224-231, January 1, 2013, the following information on semen hyperviscosity (SHV) treatment can be found:
“Semen hyperviscosity can be treated successfully in vitro by traditional methods as well as by recently developed methods. Hyperviscous semen is commonly diluted or drawn into a hypodermic needle and forced through in order to overcome the elevated viscosity, although these methods are unlikely to be completely effective because SHV is not completely a mechanical phenomenon. More direct treatments of patients presenting with SHV include over hydration, prostatic massage, and the use of parenteral hyaluronidase (injection – my note). These methods showed limited success and have proven not to be too effective.

Less dated methods used in sperm preparation for ART procedures have been shown to improve semen and sperm parameters of infertile males. Honea et al. found that the use of limited proteolysis by using α-chymotrypsin in the treatment of SHV was effective for an in vitro setting such as IVF or IUI. Zavos et al. more recently reported that limited proteolysis by α-chymotrypsin was effective in improving the use and handling of hyperviscous semen samples, and that treatment with α-chymotrypsin assisted in the recovery of high quality sperm in greater numbers than in hyperviscous samples not treated with α-chymotrypsin.”  

Conclusions:

• Though hyaluronidase is present in semen and used in both IVF procedures and for treatment of patients (via injection), it is not recommended for semen hyperviscosity treatment in vitro because there is no evidence of effectiveness.
• α-chymotrypsin was found to be an effective method for SHV in vitro treatment based on the methods listed above and years of clinical practice. 
• The QwikCheck Liquefaction Kit (based on α-chymotrypsin) product insert is enclosed, demonstrating the effectiveness of the product in clinical studies.  

DOWNLOAD QWIKCHECK LIQUEFACTION KIT PRODUCT INSERT

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It is important to understand the relationship and difference between Precision and Accuracy.  Both are important, and both showcase different aspects of a Validation or Comparison study. The image below provides a very nice visual representation:

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Question:

In addition to the obvious issues with sample loading in the SQA testing capillary, would concentration readings be higher or lower in a sample which has not fully liquefied?

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Based on the required semen sample testing volume of 0.5 mL, it is recommended to fill the entire testing capillary with a semen sample without any dilution if the ejaculate volume is >= 0.5 mL. Sometimes though the ejaculate volume may be less than 0.5 mL. The dilution mode of the SQA-V and Vision can be used in this case as follows: If the semen sample volume is >= 0.3 mL, dilute the sample using the QwikCheck Dilution media two times (1:2 or 1+1). For example, 0.3 mL of semen sample is mixed with 0.3 mL of diluent resulting in a volume of 0.6 mL. This will provide an adequate sample volume to aspirate into the SQA testing capillary and the SQA software automatically compensates for the dilution and will provide a full report, no calculations are required by the operator.

RECOMMENDATIONS
Based on the sample volume, M.E.S. recommends the following approach to testing the sample:
• Semen Volume >= 0.5 mL - No dilution is required
• Semen Volume >= 0.3 mL and < 0.5 mL - Dilution 1:2 (1+1) using the QwikCheck Dilution kit
• Semen Volume < 0.3 mL - Manual assessment [microscope or Vision Manual Mode or Vision Semi-automated Mode (10 µL automated testing + 10 µL manual motility assessment)]

REFERENCES
• WHO laboratory manual for the examination and processing of human semen - 5th ed., WHO 2010.
• SQA-V GOLD User Guide Version 2.60 I-Button
• SQA-Vision User Guide Version 104.13.2

DOWNLOAD THE FULL TECHNICAL BULLETIN HERE

SEE ALL MES TECHNICAL BULLETINS HERE

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TECHNICAL BULLETIN:  TESTING CAP PROFICIENCY SAMPLES ON SQA SEMEN ANALYZERS

For All SQA-V and SQA-VISION Systems Version 2.45 and above | UPDATE: April 27th, 2016

BACKGROUND:

The College of American Pathologists (www.cap.org) provides a nationwide proficiency challenge for automated methods of semen analysis.  These results are peer reviewed against other SQA users providing an objective and efficient way to maintain proficiency.  MES recommends this survey as an unbiased appraisal of user proficiency and system performance. 

TESTING INSTRUCTIONS FOR SQA-V/SQA-V GOLD/SPERMALITE SYSTEMS:

NOTE: The CAP proficiency material should first be run as Stabilized Sperm in the “Control” mode and re-run in the Test New Patient “Fresh” Mode if a result of 0.0 is reported in Stabilized Sperm Control mode.

SETTING UP THE DEFAULTS:

• From the main menu of the SQA-V/SQA-V Gold/Spermalite select: SERVICE > SERVICE DATA.
• From the V-Sperm computer, navigate to SET UP > SQA-V > SQA-V DEFAULTS and click “CONTINUE”.  You will now see a set-up screen displayed on the PC.
• On the bottom half of the setup screen, select the “STABILIZED SPERM” option and enter the following for Level 1 and Level 2:    
  • Lot Number = Your Sample Reference # | Target = 50 | Range = 50 | Date = Today’s date.
  • Click “APPLY” and wait a few moments for the information to transfer to the SQA-V.
  • NOTE: Do not forget to change your defaults back to Latex Beads after running your CAP sample!

TESTING YOUR PROFICIENCY SAMPLES ON THE SQA-V/SQA-V Gold/Spermalite:

1. From the MAIN MENU of the SQA select: RUN CONTROLS > LEVEL 1 and allow system to calibrate.
2. Mix the sample well by aspirating it in and out 10 times with a transfer pipette.  Avoid bubbles!
3. Load the SQA-V testing capillary and check CLOSELY for bubbles then wipe the capillary tip clean.
4. Insert capillary into the testing chamber when prompted to do so.
5. If a sample result of 0.0 is received, re-run the sample as a FRESH sample.
6. To Re-Run sample as FRESH, select “Test New Patient” from the main menu and enter the CAP sample number as the Patient ID.
7. All other entries on the first screen may be skipped.
8. On the second screen select “FRESH” under sample type and < 1 M/mL for WBC CONC. (all other entry fields may be skipped).
9. Select YES when asked if sample is sufficient for complete testing > 0.5 M/mL.
10. Allow system to calibrate and insert the capillary when prompted.
11. Report the results received.
12. Repeat Steps 1 – 11 for the second CAP proficiency material level.

TESTING INSTRUCTIONS FOR SQA-VISION SYSTEMS:

SETTING UP THE DEFAULTS:

1. From the MAIN MENU of the VISION PC, select SETTINGS > PROIFICIENCY. 
2. Enter a SAMPLE ID, DATE, and a NOTE (if necessary).
3. Click SAVE.

TESTING YOUR PROFICIENCY SAMPLES ON THE SQA-VISION:

1. Select QC / PROFICIENCY from the main MENU and touch TEST NOW on the desired option.
2. Mix the sample well by aspirating it in and out 10 times with a transfer pipette.  Avoid bubbles!
3. Load the SQA-V testing capillary and check CLOSELY for bubbles then wipe the capillary tip clean.
4. Insert capillary into the testing chamber when prompted to do so.
5. Report the results received.
6. Repeat steps 1 through 5 for all samples.

DOWNLOAD THESE INSTRUCTIONS HERE

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Click the image below for a screen shot overview of the SQA-VISION instrument.  The document may be used as a training reference tool for MES distributors and end users looking to train new staff members.  Remember, it ALL Started with a Sperm!

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Answer: Leukocytes, predominantly polymorphonuclear leukocytes (PMN, neutrophils), are present in most human ejaculates (Tomlinson et al., 1993; Johanisson et al., 2000). They can sometimes be differentiated from spermatids and spermatocytes in a semen smear stained with the Papanicolaou procedure. Differentiation is based on differences in staining coloration, and on nuclear size and shape (Johanisson et al., 2000). Polymorphonuclear leukocytes can easily be confused morphologically with multinucleated spermatids, but stain a bluish colour, in contrast to the more pinkish colour of spermatids (Johanisson et al., 2000). Nuclear size may also help identification: monocyte nuclei exhibit a wide variation in size, from approximately 7 µm for lymphocytes to over 15 µm for macrophages. These sizes are only guidelines, since degeneration and division affect the size of the nucleus.  There are several other techniques for quantifying the leukocyte population in semen. As peroxidase-positive granulocytes are the predominant form of leukocytes in semen, routine assay of peroxidase activity is useful as an initial screening technique (Wolff, 1995; Johanisson et al., 2000) / (WHO 5th ed. manual, p. 102).

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