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Research Microscopy and Histology Core Sample Submission Guidelines

Explore the Research Microscopy and Histology Core (RMHC) sample submission guidelines outlined below. RMHC cannot guarantee optimal outcomes for samples submitted outside of these specifications. 

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Formalin Fixed Paraffin Embedded Tissue 

  • Submit samples exclusively in 10% formalin; specimens stored in alcohol are not accepted.  
  • Maintain an appropriate volume (at least 20:1). 
  • Cut to no more than 3mm thick; must fit tissue cassette. 
  • Submit oriented in cassettes for optimal results. 
  • Adhere to assay-specific timing guidelines (IF/IHC and RNAscope). 

Frozen Sections 

  • Submit samples in sucrose (30% w/v for neural tissues, 20% for all others). 
  • Perform 4% paraformaldehyde perfusion, optional post-perfusion 4% paraformaldehyde fixation, and mandatory graded sucrose incubations. 
  • Dry tissue, then quick-freeze in molds using dry ice or liquid-nitrogen-cooled 2-methyl butane. 
  • See staff for protocols regarding specific skeletal muscle and eye tissues.  

IF/IHC 

  • Consult staff about optimal tissue processing, which is often antibody-dependent. 
  • Limit exposure to aldehyde-based fixatives to no more than 32 hours. 
  • Submit samples within 24 hours of harvest (Monday-Thursday only). 
  • Note: Results are not guaranteed due to commercial antibody quality variance.  

RNAscope 

  • Submit fixed/fixed-frozen samples in 10% formalin or 4% paraformaldehyde within 24 hours of harvest. Max 32 hours in fixative.  
  • Provide type of fixative and date and time fixation began.  
  • Optional: Collect fresh-frozen samples from animals perfused with 4% paraformaldehyde. Snap-freeze in liquid-nitrogen-cooled 2-methyl butane and wrap in foil.  

Plastic Sections (TEM) 

  • Consult staff for the appropriate tissue fixative for your tissue.  
  • Submit tissue as a cube no greater than 1 mm in each dimension. 
  • Submit within one week of harvest.  
  • See staff for orientation instructions. 

GeoMX DSP 

  • Use Leica Apex Superior Adhesive slides or Fisher SuperFrost Plus slides. 
  • Section slides according to Nanostring’s recommendations; tissue should be within the designated area. 
  • Prepare slides no more than one week before desired staining time.  
  • Provide a plan for ROI selection or schedule staff-assisted selection.  
  • Due to logistical complexity, schedule a project kickoff meeting with staff.