Support & Resources


Tech Tips and Protocols

Practical advice and lab tips from Biotium scientists on using fluorescence in your research

Protocols Library

A collection of protocols for immunofluorescence, western blotting, protein labeling, cell staining, and more.

Tech Tip: Multi-Color Fluorescence Imaging Using Biotium’s Tyramide Amplification Kits

Tyramide signal amplification (TSA) is an immunofluorescence technique that uses enzymatic coupling to label targets with large amounts of dye, resulting in dramatically higher signal. Here, we provide protocols for performing multiple TSA reactions on the same sample.

Tech Tip: Combining Lipophilic Membrane Dyes with Immunofluorescence

A protocol for staining fixed and permeabilized cells with CellBrite™ Cytoplasmic Membrane Dyes for immunofluorescence staining.

Tech Tip: Five steps to successful cell membrane staining & imaging

Tips for getting great staining with CellBrite™ and MemBrite™ membrane and cell surface stains.

Tech Tip: Cell Surface Stains for Live & Fixed Cells

Find the right membrane or cell surface stain for your experiment.

Tech Tip: Eight things to keep in mind for optimal gel staining with GelRed® and GelGreen®

Getting the best results with GelRed® and GelGreen® DNA gels.

Tech Tip: Using ViaFluor® SE Stains for Cell Tracing and Co-Culture

Learn how our ViaFluor® SE dyes can be used to trace cell morphology and track cells in co-culture.

Tech Tip: Imaging Bacteria Using Agarose Pads

Bacteria can be difficult to image because they are small, usually non-adherent, and often motile. Here we describe how to immobilize live or fixed bacteria using agarose pads for multi-color imaging experiments.

Tech Tip: Battling Tissue Autofluorescence

Autofluorescence in tissues can make it all but impossible to distinguish specific fluorescence staining from non-specific background. See our protocols for quenching autofluorescence with TrueBlack®.

Tech Tip: Avoiding Artifacts from UV Photoconversion of DAPI and Hoechst

Many researchers are unaware that photoconversion of DAPI and Hoechst can cause nuclear stains to fluoresce in the FITC and Cy®3 channels. Learn about the different ways to avoid DAPI photoconversion.

Tech Tip: Combined Direct and Indirect IF Using Primary Antibodies from the Same Host

Protocol for combining fluorescent secondary antibody staining with directly labeled primary antibodies, for added flexibility in multicolor staining.

Protocol: Immunofluorescence Staining of Cells for Microscopy

Our basic protocol for IF staining of adherent cells.

Protocol: Cell Surface Antibody Staining for Flow Cytometry

Basic protocol for cell surface staining for flow cytometry.

Protocol: Intracellular Antibody Staining for Flow Cytometry

Basic protocol for intracellular staining for flow cytometry.

Troubleshooting Tips for Fluorescence Staining

Common problems and solutions for IF staining for microscopy, flow cytometry, or western.

Protocol: Fluorescent Western Blotting

A basic protocol for fluorescent western detection.

Protocol: DNA Probe Labeling by PCR

Synthesize fluorescent DNA probes by PCR with our CF® Dye dUTP or dCTP nucleotides.

Protocol: Staining Cells with Hoechst or DAPI Nuclear Stains

Protocols for staining live or fixed cells with the popular blue nuclear stains Hoechst and DAPI.

Protocol: Succinimidyl Ester Labeling of Protein Amines

Protocol for labeling proteins on free amines with CF® Dye SE (NHS esters).

Protocol: Maleimide Labeling of Protein Thiols

Protocol for labeling proteins on free thiols with CF® Dye Maleimides.