Computer Aided Semen Analysis -

CASAPresented by

Vijayakumar C NChief Embryologist

Embryology Academy for Research and

Training26/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.

Thank You…