Webinars

Although the causes of non-specific background staining in immunohistochemistry (IHC) and immunofluorescence (IF) applications may differ slightly, they can obscure the detection of your specific signal. Learn about the sources of background staining and how to troubleshoot unwanted background in IHC and IF applications. 
 
Topics of discussion include: 
- Why background staining is problematic 
- How to identify sources of background 
- Strategies used to eliminate background 

If you’re in a hurry, time references have been added to help you skip to sections of interest from this overview:

Introduction 1.11 

What is background? 1.30  

Why is background in IHC & IF an issue? 3.19  

How to trouble shoot IHC & IF 4.20 

Background sources from tissue  4.45 

IHC workflow 6.11 

Identifying the source of background 7.12 

HRP Quenching methods 8.30 

AP Quenching methods 9.11 

2nd Deletion Control 9.50 

Species cross reactivity caused background 11.01 

Inadequate blocking for detection reagents 13.19 

The importance of buffer washes 14.26 

Other contributing factors 15.13 

Second deletion control for ABC methods 15.42 

Checking the complete staining system 16.25 

Immunofluorescence troubleshooting 18.20 

Tissue autofluorescence 19.42 

Methods of blocking autofluorescence 20.29 

Lipofuscin based autofluorescence 21.00 

Further tools & Resources  22.30 

Why to use them and what is an appropriate control?

View this webinar for a discussion on the types of controls to use, and what makes an appropriate positive and negative control. 
 
Topics of discussion include: 
- Controls to help troubleshoot background 
- Positive controls to assess assay sensitivity 
- Negative controls to identify false positive staining 

If you’re in a hurry, time references have been added to help you skip to sections of interest from this overview:

Introduction 1.02

Types of control 1.28

Why use controls 2.27

Positive control sections 3.30

Why use positive controls 5.20

Considerations - What to use as a positive control 8.25

Missteps in applying a positive control 10.35

Negative control sections 13.49

Omitting primary antibodies 14.24

Appropriate negative controls 15.56

Summary 22.19

Q & A session 23.04

Low signal and loss of signal frustrate investigators as these issues are often encountered when performing immunofluorescence (IF) assays. Learn techniques and strategies that maximize signal intensity and retention in cell- and tissue-based IF. 
 
Topics of discussion include: 
- Optimizing IF workflow 
- Detection methodologies 
- Preserving the stained specimen 

If you’re in a hurry, time references have been added to help you skip to sections of interest from this overview:

Introduction 0.43

Scope of webinar 1.30

Fluorophores 2.49

Tissue preparation 3.47

Positive & negative controls in the workflow 6.15

IF detection methods 7.15

Fluorophore selection 10.31

Hardware 11.38

Photobleaching 14.00

Fluorophore selection summary 15.33

Preservation of signal 16.40

Counterstains are often applied haphazardly at the end of an IHC procedure with little thought given to optimizing conditions for providing the best contrast with the specific antigen staining. Poor application or selection of a counterstain frequently causes a multitude of issues including color incompatibility, loss of substrate precipitate and obstruction of target antigen staining. Please join our webinar to learn about considerations when applying a counterstain and tips to avoid commonly encountered issues.

If you’re in a hurry, time references have been added to help you skip to sections of interest from this overview:

Introduction 0.41

Why counterstaining is important 1.00

Scope of Webinar 2.12

Counterstain Basics 2.45

Counterstain Choices 5.15

Counterstain Considerations 8.03

• Do you need one? 8.05
• Colour combinations 10.23
• Strength 11.46
• Formulation 14.48
• Specimen Fixation & thickness 16.01
• Tissue Type 18.51
• Method of application 19.48
• Shelf life 21.20

Summary 21.35

Poster offer 22.40  Request direct from 2B Scientific today: customersupport@2bscientific.com 

Q&A session 23.20

While the IHC workflow is well established, the detection reagents utilized will dictate assay parameters such as specificity, sensitivity, assay time, costs and clarity of imaging. Gain insights into selecting the most appropriate reagents for your given IHC assay in this webinar.

Introduction 0.47

How to simplify IHC 1.48

Key questions to determine which detection reagents are required 3.09

Overview of Polymer detection method 5.30

‘IHC in a box’ kits discussion 7.50

Polymer detection sensitivity 9.08

Changing existing assays 10.27

Increasing sensitivity 12.25

Changing tissue species/ tissue types 16.35

Changing Primary Antibody 18.37

Multiple Antigen Labelling 20.54

Selecting an Enzyme Substrate 23.01

Cost considerations 25.37

Q & A session 27.05

Wednesday 9th June, 2021

How to Make Excellent Antibodies for Histopathology

3.03 Overview

4.40 Introduction to Synaptic Systems and the HistoSure™ Range

7.17 Development and validation

8.40 Quality guidelines

9.45 Reference protocols

11.08 Examples of HistoSure™ staining

14.58 Humanized mice as innovative cancer models

16.22 Antibodies for human-mouse chimera

17.33 Multiplex IHC-P for validation

18.52 Recombinant antibody technology – facilitating multiplexing

Multiple Antigen Labelling for IHC

21.33Overview

23.47 Simplified double labelling

24.14 Considerations for multiple labelling

26.27 Species cross reactivity

29.05 Enzyme detection systems and substrates

33.50 Controls for multiple antigen labelling

45.08 Counterstains in multiple labelling – Do you need one?

53.42 Co-localization in multiple antigen labelling

55.19 Summary

26.46 Resources

57.30 The 2BScientific Team & Q & A session