Slide Mount with Far-red Counterstain (Cat#032347)

Aqueous Antifade Mountant with Far-red Counterstain
Laboratory Use Only, Store at -20ºC to 25ºC

The Slide Mount ^TM  with Far-red Counterstain is a water-based antifade mountant that can stably preserve fluorescence in paraformaldehyde fixed brain sections. The mountant contains TO-PRO TM -3 (T3605, Invitrogen™) that is a nucleic acid stain with far-red fluorescence similar to Alexa Fluor® 647 or Cy®5 dyes. To-Pro in the mountant is ultrasensitive to double-stranded nuclei acids (dsDNA) and can be excited by 633–647 nm lasers. This mounting medium can be used as a nuclear counterstain in brain sections labeled with commonly used green and red fluorophores. It contains antifade reagents that protect against fading or photobleaching of fluorescent proteins and dyes. It can be used in conjunction with GFP/YFP, tdTomato, Cy3, and fluorescent dye tracers, such as DiI and DiA. The mountant is supplied ready-to-use and can be stored at room temperature (RT), in a refrigerator, or in a freezer.

Slide MountTM  with Far-red counterstain was applied to brain sections. (A) 4% PFA-fixed cryostat brain section (20 µm thick) was mounted on a gelatin-coated slide and embedded in the mountant with TO-PROTM-3. Section was stored at RT for ~1 hour and then imaged using a confocal microscope. The fluorescence (magenta) was excited by a 633 nm laser and filtered through an emission channel of BP 650–710 nm. (B) Section was incubated in a primary antibody, followed by Alexa Fluor™ 543 (red) secondary antibody. Section was then embedded in the mountant, stored at RT for one week, and imaged (Red: a 543 nm laser and a ‘BP 560–615 nm’ filter; Magenta/Far-red: a 633 nm laser and a ‘BP 650–710 nm’ filter). (C,D) Sections from a GFP-transgenic mouse were embedded in the mountant, stored at RT for 3 months, and imaged. Green: a 488 nm laser and a ‘BP 500–530 nm’ filter; Magenta/Far-red: a 633 nm laser and a ‘BP 650–710 nm’ filter.

Reagent Provided:

Slide Mount^TM with Far-red Counterstain: 10 ml of solution in a dropper bottle.

The mountant is formulated for preserving fluorescence labeling in brain sections with an ultra-low background. It contains antifade reagents that suppress fading or photobleaching of commonly used fluorophores. Premixed To-Pro leads to far-red-fluorescent nuclear and chromosome counterstain with lower background.

Product Features:

• Compatible with commonly used fluorophores, providing anti-fading and anti-photobleaching.
• Premixed To-Pro leading to nuclear counterstain with far-red fluorescence.
• View and image after embedding regardless of hardening (Excitation: 633–647 nm; Emission: 650–710 nm).
• Long-term storage of coverslipped sections without needing to seal coverslip with nail polish or plastic sealant.
• Easily remove hardened mountant using PBS-T or dH 2 O.

Storage and Stability:

• Can be stored at -20°C to 25°C. Keep bottle tightly sealed and avoid strong direct light.
• Stable for six months at RT (22°C) or one year in a refrigerator (4°C).

Warranty: 12 months from the date of purchase.
Return Policy: Bioenno Tech’s return policy for this product is 90 days from the date of purchase.
Free Technical Support: Email your questions to contact@bioenno.com

Instructions for Use:

• If the Slide Mount^TM with Far-red Counterstain is stored in a freezer, thaw prior to use.
• If the Slide Mount^TM with Far-red Counterstain is stored in a refrigerator (4°C) or at room temperature (RT), use directly.

1) After fluorescent dye tracing, immunofluorescent staining (IF), or fluorescence in situ hybridization (FISH), mount brain sections upon adhesive microscope slides, such as gelatin-coated slides. Air-dry the sections/slides at RT.

Optional: Wash the dried slides in 0.01 M PBS containing 0.3% Triton X-100 (PBS-T) for 1-5 minutes and then quickly rinse (1-3 seconds) the slides with distilled water (dH₂O). Remove excess water around sections and air-dry the slides for 1-5 minutes at RT. (Please make sure the mounted sections are dried before rinsing with dH₂O.)

2) Place the slides on a level surface, apply the mountant to sections, and coverslip the sections.

• One to several drops of mountant may be needed, depending on number of sections.
• Apply coverslip slowly with the aid of surgical forceps to avoid generating bubbles.
• Touch the slide with one edge of the coverslip and slowly lower the coverslip onto the slide.
• Remove extra mountant carefully around edges by capillary action with filter paper or Kimwipes.
• No need to seal the perimeter of the coverslip with nail polish or plastic sealant.

3) Place the coverslipped slides on a level surface to harden, which may take hours or longer at RT.

• Slides can be viewed and imaged after being coverslipped. There is no need to wait for hardening.
• Mountant can be removed by immersing the slides in PBS-T or dH₂

 4) The slides can be stored at RT in a slide box or appropriate slide container. For best results, store slides in the dark.

• The slides can also be stored at 4°C, but storing at RT (22°C) works perfectly.
• Drying or long-term storage may cause air bubbles; simply remove coverslip and hardened mountant by immersing the slides in PBS-T or dH₂O, and then re-coverslipping with the mountant.

Safety and Handling Precautions:

The mountant is designed for in vitro research use only and not for drug, diagnostic, or other uses.

The mountant contains reagents that may be harmful upon contact with skin, inhalation, or ingestion. Do not pipette by mouth. Use standard precautions to avoid inhalation or contact with skin and eyes. In case of contact, wash immediately with generous amounts of water and seek medical advice. If swallowed, wash out mouth with water and immediately call a physician.

Perform experiment under a chemical hood. Wear suitable protective clothing, gloves, and eye/face protection. Wash hands thoroughly after performing the experiment.

Material safety data sheet (MSDS) is available upon request.