IMSI - Intra-cytoplasmic Morphologically Selected Sperm Injection - Stephen Harbottle imsi workshop presentation march 2011

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MRMC 2011

IMSI Workshop; use of the RI System

Stephen J. HarbottleLead Clinical Embryologist

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Introduction

• ICSI is a well established technique– First pregnancy reported in 1992

• IMSI is a natural extension to ICSI– MSOME to actively select sperm for injection– Use of high power microscopy and digital imaging

• Optimise procedure to minimise environmental stress• Identify appropriate IMSI patient group

– Maximise success rate

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MSOME

• Motile Sperm Organelle Morphology Examination (MSOME) is aimed to define the state of 6 sub cellular organelles;– Acrosome, Post Acrosomal lamina, Neck, Mitochondria, Tail, nucleus

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Defining the ‘Normal Sperm’

• Criteria for Morphologically Normal Nucleus– Smooth, symmetric, and oval configuration

• Shape– Average length 4.75 ± 0.28 μm– Long > 5.31 μm | Short<4.19 μm– Average width 3.28 ±0.20 μm– Narrow < 2.88 μm | Wide >3.68 μm

• Content– No extrusion or invagination of the nuclear chromatin mass– No vacuoles with diameter greater than 0.78 ±0.18 μm

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Significance of Nuclear Normalcy

• Normalcy of the sperm NUCLEUS was found to be a significant factor in pregnancy occurrence by ICSI procedure

Bartoov , B et al. Fertility and Sterility. 2003 Dec;80(6)

IMSI ICSI

Pregnancy Rate (%) 50% 25%

Abortion Rate (%) 13% 33%

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Examples of Vacuolation

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I No Vacuolation

II <2 Small Vacuoles

III>2 small or >=1 large

Vaculoes

IVLarge Vacuole and

other head abnormalities

Vanderzwalmen et al, 17(5): 617-627, 2008

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IMSI - Technical Development

• Intracytoplasmic Morphologically Selected Sperm Injection (IMSI)

• Clinical use first reported in 20011

– Oil Immersion x150 Lens1, DIC Illumination1

– Optimum magnification for IMSI; x6000 - x70001,2,3

• Use of oil and DIC microscope not optimal for clinical practice– Alternative oil free digital system developed to improve IMSI

efficiency without compromising performance & results 1 Souza Setti, A et al. Reprod Biomed Online. 2010 Oct;21(4):450-5

2 Nadalini, M et al. Reprod Biomed Online. 2009;19 Suppl 3:45-55

3 Ai, L et al. Zhonghua Nan Ke Xue. 2010 Sep;16(9):826-9

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IMSI – The RI System Hardware

IMSI Workstation Digital Camera Objective

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IMSI UK Update – Study #1

• Patient Group– ICSI Failure (>3 cycles)– Repeated miscarriage– Teratozoospermia (<4% Normal Forms)– Poor blastocyst development

• RI IMSI system– Magnification x6300 (x60 air objective with digital zoom)

• Methodology– All sperm selected and collected before oocyte injection

commenced

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Results

Chau et al, Fertility 2011

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IMSI UK Update – Study #2

• Patient Group– Patients with more than 6 eggs had ICSI & IMSI

• Sperm graded prior to injection;

• RI IMSI system– Magnification x6300 (x60 air objective with digital zoom)

Grading Category Description

Grade I No Vacuolation

Grade II <2 Small Vacuoles

Grade III >2 small or >=1 large Vaculoes

Grade IV Large Vacuole and other head abnormalitiesVanderzwalmen et al, 2008

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Results

Best et al, Fertility 2011

(n=114) (n=108)

Grade Description

I No Vacuolation

II <2 Small Vacuoles

III>2 small or >=1 large

Vaculoes

IVLarge Vacuole and other

head abnormalities

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MRMC 2011

IMSI – Our Plans

• Clinical IMSI to roll out in June 2011• Partnership with Research Instruments

– RI system has been demonstrated to be the most user friendly, cost and time effective system for clinical use

– Randomised prospective treatment study to contribute to UK IMSI dataset

– Prospective clinical study to assess the effect of sperm preparation technique

– Further work on sperm quality (grade) and its correlation to rates of fertilisation, cleavage, embryo development & implantation

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Optimising the IMSI procedure

• Use PVP to slow sperm and aid analysis and capture

PVP MSOME Selection (5µl)

PVP Injection (5µl)

Oocyte Injection (5µl)

• Use of Micropipette to trap a single good prognosis sperm for detailed analysis prior to selection

• Collect and store good prognosis sperm prior to loading oocytes into the injection dish

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The Isolator ConceptThe Isolator Concept

• Isolator units linked together with ‘pass through’ incubators linking units together

• Workspace heated, gassed and humidified to user specification

• Built in HEPA and carbon filtration system‒ Effective against particulates and VOC’s

• Gametes and embryos are never exposed to suboptimal conditions

• Significant increase in clinical treatment results

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Summary

• MSOME is a valuable clinical tool in the assessment of male factor infertility

• IMSI is an effective treatment option for patients following failed ICSI treatment

• IMSI may confer some protection against miscarriage when compared to ICSI

• IMSI may improve success in cases of unexplained infertility

• Optimise IMSI technique to reduce gamete stress• Develop systems to maintain biological conditions

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Thank you for your Attention

Terima Kasih