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• New cleaning kits will include the updated instructions and will have an outside label indicating a new kit.
UPDATED CLEANING KIT INSERT (click to download):
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Background:
The need for external quality control schemes for confirming diagnostic procedures/equipment is of utmost importance. A number of multicentre studies on semen quality confirm this fact (1, 2). Quality control materials are best suited when they closely replicate the actual material being tested. However, due to the biological nature of semen, providing external quality control materials presents a challenge. Semen is motile for a relatively short period of time and the sample degenerates quickly. Semen samples prepared by proficiency/quality control organizations are based on adding a fixative media to the donor semen samples. As a result, the original nature of the fresh semen is distorted. In addition, there can be significant batch-to-batch variations in the proficiency challenge sample matrix. This causes problems running such samples in the SQA-V - especially at the low end of concentration.
Matrix and Spermatozoa Modification by Fixative Materials
Stabilized sperm is made from a fresh semen sample. This fixative process impacts the original semen sample as described below:
• The spermatozoa are shrunk because of fixation which causes a change to their size and shape.
• The seminal plasma is diluted with a fixative or completely replaced with a media causing a decrease in the sample viscosity. The SQA-V algorithm works based on the optical density of SEMINAL PLASMA (not the dilution media or modified “seminal plasma” used in stabilized sperm).
• A decrease in sample viscosity and density results in rapid cell sedimentation and uneven distribution of the cells throughout the volume of the sample. These factors may result in inconsistent concentration of cells in the samples received by the labs and in the aliquots taken for analysis. The sample tested may no longer reflect the target value established by the manufacturer.
All of these facts will adversely impact the accuracy of automated testing especially in low concentration samples because the prepared quality control material is not the same as a normal semen sample. The SQA-V is designed to run semen samples within one hour of collection and the concentration dynamic range of the SQA-V is 2 M/ml on the low end. Therefore, running a quality control sample of very low quality on the SQA-V, may result in a ZERO reading.
Running Low Level Stabilized Sperm Samples on the SQA-V GOLD System
If a stabilized sperm sample run on the SQA-V GOLD in the stabilized sperm Quality Control mode results in ZERO (beyond 2 M/ml dynamic range), re-run the sample in the Fresh mode as outlined below:
· Turn on the SQA-V GOLD system and wait until auto-calibration/self-testing is completed.
· Go to: MAIN MENU>TEST NEW PATIENT and enter:
o PATIENT ID: Sample #
o BIRTH DATE: Test date
· From the next screen select:
o SAMPLE TYPE: FRESH
o WBC CONC: < 1 M/ml
o Select “Yes” when asked “IS SAMPLE VOLUME SUFFICIENT FOR COMPLETE RESTING >= .5 ml?”
· Vortex the stabilized sperm sample.
· Transfer the sample from the original vial to the 10-ml collection cup and mark it with the sample #.
· Mix the sample thoroughly and immediately fill the SQA-V capillary and run the test.
References
1. WHO laboratory manual for the examination and processing of human semen - 5th ed., World Health Organization, 2010.2. Auger J. et al. Intra- and inter-individual variability in human sperm concentration, motility and vitality assessment during a workshop involving ten laboratories, Human Reproduction, 2000, 15, 11, pp. 2360-2368.
DOWNLOAD THIS TECHNICAL BULLETIN HERE
Direct Link to this Post: Permalink • Standard #2: 100 micron depth (haemocytometers) that require sample dilution.
Please contact MES directly for additional questions about the type of chamber you use, and Remember, it ALL Started with a Sperm!
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This document includes pages 67 through 95 of the WHO Laboratory Manual for the Examination and Processing of Human Semen FIFTH EDITION. This updated version of the WHO manual does a very good job of showcasing normal vs. abnormal forms including all relevant defects. Dr. Thinus Kruger assessed the slides presented personally and in addition to grading them provides comments to further explain his judgements. It is highly recommended that all SQA-V users, and facilities conducting manual semen analysis become familiar with this new criteria as it is fast becoming the world standard. The document also provides 12 plates worth of numbered cells which can be assessed by the operator for training and proficiency purposes (don't look at the answers ahead of time!) The document can be downloaded by clicking the image above or by clicking HERE
Remember, it ALL Started with a Sperm.
Direct Link to this Post: Permalink Maintaining an accurate backup method for any automated analyzer in the laboratory is important. When it comes to semen analysis, there are many different counting chamber options available. Correlating manual methods to automated methods can be challenging, especially when it comes to semen analysis. The first step is picking the correct tool for the job. Below are the WHO 5th Edition Manual's recommendations for sperm concentration counting chambers. To achieve the best possible correlation to the SQA-V Gold Automated Sperm Quality Analyzer, and maintain the most accurate backup method possible these recommendations should be followed. In particular please note that disposable plastic counting chambers can lead to increased variability.
2.7.1 Types of counting chambers (From Page 34 of the WHO 5th Edition Manual for Semen Analysis)
Direct Link to this Post: Permalink Thursday, February 6, 2014
BACKGROUND:
The SQA-V technology for assessing sperm concentration is based on the principle of spectrophotometry coupled with software filters that eliminate ‘background’ factors that can influence the SQA-V results.
OVERVEIW:
It is well known that a semen ejaculate contains cells other than spermatozoa (1). These include epithelial cells from the genitourinary tract, as well as leukocytes (WBC) and immature germ cells (2). The presence of these non-sperm cells in semen may be indicative of testicular damage (immature germ cells), pathology of the efferent ducts (ciliary tufts) or inflammation of the accessory glands (leukocytes) (1).
These non-sperm cells or along with seminal plasma are all considered ‘background’ factors that interfere with the accurate assessment of sperm concentration using spectrophotometry technology. When the SQA-V assesses sperm concentration, the ‘background’ is subtracted by the software in order to eliminate its impact on the final and accurate assessment of sperm concentration. The normal SQA algorithm compensates for ‘normal’ levels of ‘background’ which are seen in most semen, however, abnormal levels of ‘background’ require a different algorithm that compensates for this. Hence, the operator is asked to assess for abnormal levels of WBC (the most contributing background factor) and select ≥ 1 M/ml in order to activate the appropriate algorithm required to measure the sample’s concentration.
The normal value for semen WBC is < 1 M/ml. In cases of inflammation, the leukocyte level is ≥ 1 M/ml and the semen will also contain seminal plasma compounds associated with inflammation. This increases the level of ‘background’ to subtract from the algorithm. This is why it is important to test if WBC’s are NORMAL or ABNORMAL and to select the proper input on the PATIENT DATA ENTRY SCREEN in order to activate the appropriate algorithm for sample testing.
INSTRUCTIONS:
WBC levels in semen can be tested using one of the following procedures:
• Using QwikCheck™Test Strips - recommended (please refer to the SQA-V User Guide Version 2.60 I-Button, Appendix 7: Measuring WBC's in Semen).
• Using the SQA-V visualization screen or the V-Sperm and visually/manually assessing the sample for WBC’s (please refer to the SQA-V User Guide Version 2.60 I-Button, Appendix 4: Counting Cells using the SQA-V Visualization System and Appendix 7: Measuring WBC's in Semen).
MANUFACTURER’S RECOMMENDATION:
WBC assessment should be conducted on FRESH and WASHED semen samples and the results entered in the SQA-V PATIENT/SAMPLE DATA ENTRY screen PRIOR to running automated semen analysis on the SQA-V and before any sample treatment with chymotrypsin or any other agent (enrichment, washing, dilution, etc.).
REFERENCES:
1. WHO laboratory manual for the examination and processing of human semen - 5th ed., World Health Organization 2010.
2. Johanisson E et al. (2000). Evaluation of “round cells” in semen analysis: a comparative study. Human Reproduction Update, 6:404-412.
Dr. Lev Rabinovitch, CTO MEDICAL ELECTRONIC SYSTEMS
Distribution: All SQA Users
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Direct Link to this Post: Permalink - pH: Normal values for semen pH are generally ≥ 7.2; the pH pad reports a range of 5.0 to 8.5.
- Leukocyte Level: Normal values for semen Leukocytes are < 1 M/ml; the Leulocyte pad reports semi-quantitative results: NEG is <1M/mL and POSITIVE is ≥1 M/mL. If the Leulocyte pad color is equal to or exceeds the darkest lavender color of the chart within 60 seconds, the result is ≥1 M/mL, otherwise it is <1M/mL.
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Direct Link to this Post: Permalink BACKGROUND:
The WHO 5th edition manual provides recommendations for testing post-vasectomy semen samples.
OVERVIEW:
Per the WHO 5th edition manual, it is not recommended to centrifuge the post-vasectomy sample if the technician is searching for motile spermatozoa, as centrifugation impacts sperm motility. If no spermatozoa are found in the ejaculate it is recommended to re-assess the sample after centrifuging and re-suspending the pellet in a small volume of seminal plasma.
INSTRUCTIONS:
Fast scan:
o Prepare a standard slide per the user guide instructions (10 μl liquefied ejaculate, 22mmx22mm coverslip). Search for spermatozoa using the SQA-V visualization system, set at “Zoom Out”.
If 1-2 MOTILE spermatozoa are found in each field of view:
o Run the sample in the SQA-V Post-vasectomy mode.
If >=2 MOTILE spermatozoa are found in each field of view:
o Run the sample in the SQA-V FRESH mode.
If only IMMOTILE spermatozoa are found:
o Count the sperm cells manually using the SQA-V visualization system set at Zoom Out. Count in duplicate using the same slide. Compute the total number of sperm divided by the number of assessed fields of view. This will roughly represent the sperm concentration in millions per ml.
If NO spermatozoa are found:
o Centrifuge the semen sample at 3000g for 15 minutes.
o Decant most of the supernatant.
o Re-suspend the sperm pellet in approximately 50μl of seminal plasma.
o Test the sample manually in duplicate using a standard slide (please see above).
o The presence of spermatozoa indicates cryptozoospermia; the absence of spermatozoa indicates azoospermia.
o This method cannot be used to determine sperm concentration or total sperm number.
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