Computer Aided Semen Analysis - CASA Presented by Vijayakumar C N Chief Embryologist Embryology Academy for Research and Training 26/A Raju Industrial Estate Penkar Pada Road MIRA-401104 India Ph: +91-22-2845 6768/ 69/ 70. Fax: +91-22-2845 6766 Email: support@ivftraining.com URL: www.ivftraining.com

Introduction Inter lab and intra lab deviations in Semen Reports Inter personal deviations in Semen Analysis Reports Heisenberg's Uncertainty Principle

Requirement of CASA To ensure indifference in semen count, motility and morphology estimates To reduce inter personal variations and to standardize observables with higher limits of acceptance Reliable reproducibility Option to have databank for a patient’s records

Overview High Resolution camera attached to Microscope takes video of semen sample for a prefixed time, till 200 sperms are imaged The video is then examined through software for sperms with respect to their number, their motion characters with in the frame User feeds the data on chamber depth, volume of sample loaded The software now provides you with the exact count and motility as per WHO guidelines as m/ ml

Count and motility estimation by CASA

Morphometry Stained semen smear/ unstained sample using AAV Technology (Negative phase Contrast in 40x) is exposed to the M’scope and image is grabbed through camera 100/ 200 sperm images are grabbed Then the images are analyzed after super imposing with standard normal/ abnormal sperm templates by the software Results are expressed as percent normal/ Abnormal Capable of performing WHO/ Strict Criterion evaluations

Morphometry

DNA Fragmentation in Sperms Results as rise of ROS in the sample > ROS gives rise to Fragmented DNA Fragmented DNA of sperms is responsible for recurrent Miscarriages post ICSI Sperms with Fragmented DNA appear as normal sperms even under 1000x magnification

How to identify DNA Fragmentation Indirectly by ROS estimation in semen Directly by Tunnel/ SCSA/ fluorescent assay/ Halo sperm assay Different techniques have different cut off values Most reliable is Halo sperm technique CASA can help in inter person variation of results with rigid limits of assay parameter

DNA Fragmentation analysis by CASA

Rationale in selecting CASA Many manufacturer’s are there We need a system which can test all parameters Disposables like chambers/ slides etc., must be available from same co., All hardware and software must be available from same co.,

Which one to offer then…… Proiser ISAS Lab CASA system Proiser is a co., from Spain formed out of Researchers in Andrology Proiser has developed CASA systems for various sperm samples for decades Proiser system is complete i.e., with Microscope, Negative Phase Contrast system, Camera, heated stage, Computer and software With Proiser You can test Count, Motility, Morphology and DNA fragmentation in one software window Disposables are developed specially for Proiser and are freely available.

What is offered in Proiser ISAS system

Constituents of Proiser ISAS CASA What is ISAS? – Integrated Semen Analysis System which is offered in two varieties ISAS – Complete System (user defining is Required) ISASLAB – Complete System (ready to use)

ISAS – Complete System modules ISAS-MC - motility & concentration only ISAS-MCMf - Motility, Concentration, Morphometry/ morphology ISAS-MCMfd - Motility, Concetration, Morphometry and DNS fragmentation

ISASLAB – system Modules LAB-MC – Motility & Concentration only LAB-MCM – Motility, Concentration and Morphology (without staining, simultaneously) LAB-MCM-STM – Motility, Concentration and Morphology with stained slides No DNA fragmentation module in ISASLAB.

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