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 40 x) 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 1000 x 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.
Thank You…
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