Ventana BMKXT IHC Overview User Training Ventana BMKXT IHC Overview 2009 picture placeholder

목차 Ventana Background 1 BenchMark® XT Instrument 2 Detection kit 3 IHC Procedure on BMKXT 4 5 Protocol & Prep kit Use

A Member of the Roche Group Ventana Background A Member of the Roche Group 1985년 창립 Head Quarter : Arizona Tucson in USA IHC, ISH, Special Stains 자동화 장비 및 시약 공급 43개 나라에 8,500 여대 setting 우리나라에 현재 48 대 설치

특수한 학습자 BenchMark® XT Instrument Full Auto No Hand Fast Reliable …… Flexible

IHC, Double Staining, IF, ISH, SISH BenchMark® XT Instrument Full Automation Baking 에서부터 counter staining 까지 Flexible Automation IHC, Double Staining, IF, ISH, SISH

30 Slide per run BenchMark® XT Instrument Turn Around Time (TAT) 3시간(IHC), 4시간 (ISH), 6시간 (SISH) Through Put 60 slide/day + 30 slide (O/N) = 90 slide Individual Heater 30가지의 다른 protocol 사용가능

Reagent volume BenchMark® XT Instrument 컴퓨터에 8 unit 연결하여 사용 가능 Only 100ul : 시약 소모량, waste양 작다! 컴퓨터에 8 unit 연결하여 사용 가능 protocol, reagent 공동으로 사용

BMKXT Staining SPECIMEN

Perfect & Fast Staining! The BenchMark of Quality 학습부진 학생 Perfect & Fast Staining! Incubation time Running time 감소 Concentration requirement 감소 균일한 Staining

BenchMark Technology – Vortex Mixing Reagent Distribution : 반응시간 단축 . 조직 전체의 균일한 염색

BenchMark Technology –Buffer Jet Slide Washer Complete Washing : background 염색의 감소

BenchMark Technology –Liquid Coverslip Complete Slide Coverage : 시약의 증발 방지

BenchMark Technology –Temperature Integrated reaction temperature control : 슬라이드 전체의 균일한 온도 조절

BenchMark IHC Reagent Protease Primary Ab Detection kit (DAB) 전처리용 Enzyme Primary Ab 타회사 Ab는 Prep kit 사용 Detection kit (DAB) i view detection kit(IHC) 1.Inhibitor 2.Biotinylated Ig 3.Streptavidin HRP 4.DAB Chromogen 5.DAB H202 6.DAB copper Ultra view detection kit(IHC) 2.HRP multimer 3.DAB Chromogen 4.DAB H202 5.DAB copper Hematoxylin Counterstain Blueing reagent Hematoxylin 색을 더 밝게 나란히 나란히

BenchMark IHC Bulk solution EZ Prep LCS Reaction buffer SSC CC1 CC2 Optional 탈파라핀 Liquid Coverslip Washing buffer Ph8.0 전처리 Ph6.0 SISH * 작동 전 Bulk Volume 확인!

i view detection kit IHC BASIC – Detection LSAB secondary Ab Biotin labeled Streptavidin-HRP primary Ab antigen 발색제

Ultra view detection kit IHC BASIC – Multimer Ultra view detection kit More Sensitive! Biotin free !! Fast !! secondary Ab HRP labeled primary Ab 발색제 antigen

Polymers vs Multimer [polymer] Diffuse & high background 염색 결과 HRP P Diffuse & high background 염색 결과 [multimer] HRP Polymer 는 backbone 에 의해서 polymer detection 의 크기 와 모양의 변화로 인하여 sensitivity 에 영향을 미쳐 번지는 듯한 염색결과로 인한 high background 염색 결과를 초래할 수 있습니다 Background Staining 의 감소 !!! arm 18

Step by Step IHC Staining on BMKXT Baking paraffin wax coated slide SPECIMEN Baking 75℃ 4 minutes

Step by Step IHC Staining on BMKXT Deparaffinization and Rehydration paraffin wax coated slide SPECIMEN EZ Prep : 파라핀 제거

Step by Step IHC Staining on BMKXT Deparaffinization and Rehydration tissue section SPECIMEN EZ Prep : 파라핀 제거

Step by Step IHC Staining on BMKXT Deparaffinization and Rehydration Antigen Retrieval(CC1) 20:00 SPECIMEN CC1 : 항원 복원을 위한 전처리 과정 항체에 따라 Short 8분, mild 30분, extend 60분 선택 * 항원복원이 필요없거나 protease 처리해야하는 antibody 도 있음

3% H202 으로 내재된 peroxidase 활성 억제 Step by Step IHC Staining on BMKXT washing Inhibitor Deparaffinization and Rehydration Antigen Retrieval(CC1) inhibitor SPECIMEN Inhibitor 4 minutes 3% H202 으로 내재된 peroxidase 활성 억제

Primay Antibody incubation RT. 37C, 42 C 선택 Step by Step IHC Staining on BMKXT Primary Antibody Deparaffinization and Rehydration Antigen Retrieal (CC1) Inhibitor Rinse HER-2 Antibody SPECIMEN Primay Antibody incubation RT. 37C, 42 C 선택

Biotinylated 2차 antibody : mouse IgG+mouse IgM+ rabbit IgG Step by Step IHC Staining on BMKXT Biotin Ig Rinse Deparaffinization and Rehydration Antigen Retrieval Inhibitor Primary Antibody Biotin Ig SPECIMEN Biotin Ig 8 minutes Biotinylated 2차 antibody : mouse IgG+mouse IgM+ rabbit IgG

Streptavidin HRP 8 minutes Step by Step IHC Staining on BMKXT Deparaffinization and Rehydration Streptavidin HRP Antigen Retrieval Inhibitor Primary Antibody Biotin Ig Rinse Streptavidin HRP SPECIMEN Streptavidin HRP 8 minutes

Step by Step IHC Staining on BMKXT DAB +H202 Deparaffinization and Rehydration Antigen Retrieval Inhibitor Primary Antibody Biotin Ig Streptavidin HRP Rinse DAB SPECIMEN DAB + H202 8 minutes

DAB Color 를 radish brown color 로 Step by Step IHC Staining on BMKXT Deparaffinization and Rehydration Antigen Retrieval Inhibitor Primary Antibody Biotin Ig Streptavidin HRP DAB + H202 Rinse COPPER copper SPECIMEN COPPER 4 minutes DAB Color 를 radish brown color 로

Step by Step IHC Staining on BMKXT Deparaffinization and Rehydration Antigen Retrieval Inhibitor Primary Antibody Biotin Ig Streptavidin HRP DAB + H202 Rinse COPPER Hematoxylin Hematoxylin Hematoxylin SPECIMEN Hematoxlyin : 4분

Step by Step IHC Staining on BMKXT Deparaffinization and Rehydration Antigen Retrieval Inhibitor Primary Antibody Biotin Ig Streptavidin HRP DAB+H202 Rinse COPPER Hematoxylin Blueing reagent Blueing reagent SPECIMEN Blueing reagent : 4분 Hematoxylin 색을 좀더 파랗게

BMKXT Protocol 만들기 3 detection kit 선택

* 있는 번호는 이미 저장된 것이므로, 없는 번호 선택! 4 5 * 있는 번호는 이미 저장된 것이므로, 없는 번호 선택!

Prep kit – antibody 입력

concentration unknown 체크하면 입력안해도 됨 Prep kit – antibody 입력 concentration unknown 체크하면 입력안해도 됨 Expiration date 는 최대한 길게!(입력된 날짜가 지나면, 시약 사용 불가)

Prep kit – 입력된 antibody를 prep kit에 묶기

Prep kit – 입력된 antibody를 prep kit에 묶기

Prep kit – 입력된 antibody를 prep kit에 묶기

Prep kit 사용 유의점 ! register 한 후 사용이 가능 250T(Prep kit 1 ~ 30번), 100T(Prep kit 31번~150번) 유효기간 : register 한 후 18개월 장비에 setting 하기 전 주사기를 사용하여 압력을 생성 * prep kit 번호에 원하는 antibody 를 fill 했는지 확인 Protocol 에서 Antibody 선택시, 맞는 prepkit에 check!

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