Super-resolution microscopy encompasses a broad family of imaging techniques that push beyond the diffraction limit of traditional light microscopy, revealing the finer details of biological structures. These techniques rely on extremely precise control over the excitation, emission, and image acquisition of fluorescently labeled cells and tissues. Consequently, the quality and efficiency of super-resolution imaging relies heavily on the fluorescent properties of the probe. Recent advances in super-resolution microscopy have demonstrated the importance of high-performance fluorescent probes. Biotium’s CF® Dyes have met this demand with industry-leading brightness and photochemical switching properties over other commercially available fluorescent probes. Moreover, CF® Dyes have been published in dozens of studies for a wide range of super-resolution applications.

Superior Performance for STORM Imaging

Several CF® Dyes were developed and validated for optimal performance in Stochastic Optical Reconstruction Microscopy (STORM). This includes Biotium’s red-excited CF® Dyes such as CF®647, CF®660C, and CF®680 which display superior brightness and unique photoswitching properties that are ideal for STORM. However, while STORM imaging commonly relies on photoswitchable red-excited dyes, finding comparable dyes for other excitation sources to produce quality multi-color STORM images remains a challenge. In response, Biotium has developed several green-excited dyes developed for optimal performance in STORM. Specifically, green-excited CF®583R and CF®597R were developed in collaboration with UC Berkeley and STORM scientist Ke Xu, PhD for exceptional performance in multi-color STORM imaging (Figures 1 and 2). In addition, green-excited CF®568 has also demonstrated better performance than Cy3b (Figure 3). CF®594ST is Biotium’s unique version of CF®594 developed specifically for STORM. See below for a full list of validated CF® Dyes for STORM and available products.

CF® Dyes Validated for STED and FLIM Imaging by STELLARIS 8 STED Confocal Microscope

CF® Dyes were validated for high-quality super-resolution imaging of microtubules by Leica’s STELLARIS 8 STED FALCON platform. See the table on the right for fluorescent lifetime data for several CF® Dyes.

Don’t Just Take Our Word for It!

Check Out More Publications Validating CF® Dyes for STORM

Sign up to our e-newsletter to download our Literature Digest: CF® Dyes for STORM Super-Resolution Microscopy.

Antibody Labeling Kits

Our Mix-n-Stain™ STORM CF® Dye Antibody Labeling Kits allow you to label 50 ug of your antibody with one of Biotium’s STORM CF® Dyes to produce an antibody conjugate with a low 1-2.5 DOL (degree of labeling) optimal for super-resolution STORM (Stochastic Optical Reconstruction Microscopy). Labeling takes just 30 minutes, with minimal hands-on time and no purification. Click here to learn more about Biotium’s antibody labeling kits.

• Choice of 8 CF® Dye colors ideal for STORM
• CF®583R and CF®597R are novel green-excited STORM dyes
• Labeling optimized to provide low 1-2.5 DOL
• Less than 30 seconds of hands-on time
• 30 minutes total reaction time
• No purification, 100% recovery

Reactive CF® Dyes

Developing a custom probe for super-resolution applications? Biotium offers CF® Dyes in a wide array of reactive chemistries to suit your research needs.

• Succinimidyl esters for labeling antibodies, proteins, or other amine-containing molecules
• Maleimide and MTS CF® Dyes for labeling thiols
• Aminooxy and hydrazide reactive CF® Dyes for labeling carbonyl groups or carbohydrates
• Azides, alkynes, & BCN CF® Dyes for highly specific bioorthogonal labeling

MemBrite® Fix stains are validated for super-resolution and other specialized imaging applications

MemBrite® Fix stains are novel reactive fluorescent dye stains that react irreversibly with cell surface proteins, for staining that can withstand formaldehyde or alcohol fixation, and detergent permeabilization. Unlike lectins such as WGA, where surface staining varies between cell types, MemBrite® Fix dyes react widely with cell surface proteins for more uniform staining.

Several MemBrite® Fix dyes have been validated in super-resolution imaging applications or 2-photon microscopy. MemBrite® Fix-ST dyes are specialized dyes recommended for super-resolution imaging by STORM. MemBrite® Fix-ST dyes can also be used for standard microscopy applications; however, MemBrite® Fix dyes are generally more photostable than MemBrite® Fix-ST dyes. See the table on the right for a list of MemBrite® Fix dyes validated for super-resolution and other specialized imaging applications.

LysoView™ Dyes are fluorescent stains for imaging lysosome localization and morphology in live cells. The dyes accumulate in the low pH environment of the lysosomes, resulting in highly specific lysosomal staining without the need for a wash step. LysoView™ 540 and LysoView™ 633 exhibit pH-sensitive fluorescence, further enhancing the specificity of staining. LysoView™ 488 has been validated in super-resolution imaging by SIM (see highlighted citation below), while LysoView™ 650 fluorophore is compatible with super-resolution imaging by SIM and STED.

Validated Stains for Imaging Extracellular Vesicles by STORM

Extracellular vesicles (EVs), including exosomes, are lipid-bound vesicles that are released from cells. EVs display specific surface proteins and can carry nucleic acids and other cargo, allowing them to transfer biological information between cells in different parts of the body. Therefore, EVs are increasingly studied for their potential use in drug delivery and medical diagnostic applications.

ExoBrite™ STORM EV Membrane Stains were developed for STORM imaging of EVs and are available in four CF® Dyes validated for STORM.  ExoBrite™ 560/585 and ExoBrite™ STORM CF®647 have been validated for dSTORM on the ONI Nanoimager, allowing the study of fine morphological details and co-staining with EV biomarkers such as tetraspanin proteins CD9, CD63, and CD81.

ExoBrite™ STORM EV Membrane Stain advantages:

• Dive deeper into EV morphology and structure
• Available in 4 CF® Dyes validated for STORM imaging
• Allows antibody co-staining and localization studies with EV biomarkers