{"id":29447,"date":"2018-03-14T17:20:36","date_gmt":"2018-03-14T16:20:36","guid":{"rendered":"https:\/\/www.micropticsl.com\/sca-tricks\/"},"modified":"2026-06-29T14:42:14","modified_gmt":"2026-06-29T12:42:14","slug":"sca-tricks","status":"publish","type":"post","link":"https:\/\/www.micropticsl.com\/es\/sca-tricks\/","title":{"rendered":"SCA Tricks"},"content":{"rendered":"
<\/p>\n
\n
\n

Tricks for optimizing the analysis of sperms with the SCA<\/h2>\n

 <\/p>\n

\"\"<\/p>\n

\n

 <\/p>\n

1.<\/strong><\/span> The presence of bubbles inside the counting chamber may alter the results obtained by your SCA.<\/h3>\n

This occurs when:<\/p>\n

a) Non-optimal counting chambers are used for automatic analysis.<\/strong><\/p>\n

b) The counting chamber is not filled in correctly.<\/strong><\/p>\n

– Micropipettes must be correctly calibrated to prevent air bubbles during sample aspiration\/release.<\/p>\n

– Counting chambers must be filled in progressively and in one step<\/strong>. It\u2019s recommended to press slowly the plunger button<\/strong> of the pipette allowing a continuous and homogeneous sample release<\/strong>.<\/p>\n

 <\/p>\n

2.<\/strong><\/span> Use the Intelligent Filter.<\/h3>\n

The intelligent filter option in the \u201cConfiguration\u201d menu allows the distinction of sperms (cells with tail) from debris\/other cells that have no tail. The activation of that setting offers a higher accuracy throughout the calculation of sperm concentration, for the detection of artefacts that could be erroneously classified as sperms is omitted.<\/p>\n

NOTE<\/strong>:<\/span> When working with a USB Basler acA-1300 camera, it\u2019s recommended to change the resolution of the \u201cCapture\u201d setting to optimize the sensitiveness of the analysis:<\/p>\n

\"SCA

SCA 6.3 version: Select \u201cLow\u201d Capture Resolution<\/p><\/div>\n

 <\/p>\n

\"SCA

SCA 6.2 version: Change the Resolution from 100% to 60%<\/p><\/div>\n

 <\/p>\n

3.<\/span><\/strong> Check the \u201cComparison\u201d option for getting robust results.<\/h3>\n

The \u201cComparison\u201d graph allows to check the homogeneity of the captured fields. When fields are homogeneous, these are shown in blue colour and concentration values are similar in the graph:<\/p>\n

\"Comparison<\/p>\n

However, when fields show very different concentrations (i.e. due to the presence of aggregations or agglutinations), these are shown in red and the graph for concentration is heterogeneous:<\/p>\n

\"Comparison<\/p>\n

In those latter cases, it\u2019s recommended to delete the dissimilar fields shown in red and capture more fields (if required) to get more reliable results.<\/p>\n

 <\/p>\n

 <\/p>\n

\"\"<\/p>\n

\n

 <\/p>\n

1.<\/strong><\/span> Getting an optimal staining.<\/h3>\n

Staining protocols have been standardized considering specific laboratory environmental conditions (50% relative humidity and temperature to 21-23\u00b0C). But, since many laboratories are not working under these conditions, staining procedures must be adapted to every specific laboratory conditions before performing an automatic analysis.<\/p>\n

Intrinsic seminal factors may also hinder the automatic detection. Viscosity and seminal proteins commonly result in a dim and noisy background once semen smears are stained. In these cases, a simple sperm washing is recommended to get a clearer background and a higher contrast between the distinct parts of sperm cells.<\/p>\n

 <\/p>\n

2.<\/strong><\/span> Capturing accurately at high magnifications.<\/h3>\n

Morphological analysis at 100x but also at 60x require the use of a mounting medium (i.e., Eukitt, DPX, or similar) and a coverslip to get optimal focus and a good contrast of the different sperm head regions and vacuoles. If smears are not mounted, overestimation of the size and non-realistic results could occur.<\/p>\n

Non-mounted (left) vs Mounted (right) slide at 60x:<\/p>\n

\"Non-mounted<\/p>\n

\"\"<\/p>\n

 <\/p>\n

Nuria Medarde, PhD<\/strong>
\nBiologist
\nMICROPTIC S.L.<\/p>\n

<\/div><\/div><\/div>\n<\/div>\n

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