Difference between revisions of "Making Diluted Solutions of MicroSpheres"

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=== Procedure - From liquid with ~ 10% solids ===
 
=== Procedure - From liquid with ~ 10% solids ===
  
* Shake microsphere bottle to evenly disperse the particles.
+
* Shake primary microsphere bottle to evenly disperse the particles.
  
 
* Remove 100ul of cloudy white microsphere solution and dilute with 20ml of deionized water (200:1 dilution).
 
* Remove 100ul of cloudy white microsphere solution and dilute with 20ml of deionized water (200:1 dilution).
  
* Bovine Serum Albumin (BSA) and (for non-biological particles) 1% polyethylene glycol can be added as anti-stick agents.
+
=== Anti-Stick Agents ===
  
* For biological particles BSA and Phosphate-Buffered Saline (PBS - 0.8% NaCl, 0.02% KCL, 0.02M PO4, pH 7.3 (percentage by weight)) can be used as anti-stick agents.
+
* Bovine Serum Albumin (BSA) at 20-25mg/ml in deionized water can be applied to the glass microscope slide (and coverslip) and allowed to evaporate off (to inhibit particle sticking to the glass surfaces).
 +
 
 +
* For non-biological particles, 1% polyethylene glycol can be added to the microsphere solution as an anti-stick agent.
 +
 
 +
* For biological particles the BSA application and Phosphate-Buffered Saline (PBS) (Sigma-Aldrich P5368-10PAK) can be used as anti-stick agents.
 +
 
 +
* If particles begin to stick to the slide surfaces, a 3 minute sonication can increase the mobility of particles.
 +
 
 +
[http://en.wikipedia.org/wiki/Phosphate_buffered_saline PBS Wikipedia]
  
 
''Students: If this recipe is inadequate (too many or too few particles) please revise. ''
 
''Students: If this recipe is inadequate (too many or too few particles) please revise. ''

Latest revision as of 18:19, 4 September 2015

Procedure - From liquid with ~ 10% solids

  • Shake primary microsphere bottle to evenly disperse the particles.
  • Remove 100ul of cloudy white microsphere solution and dilute with 20ml of deionized water (200:1 dilution).

Anti-Stick Agents

  • Bovine Serum Albumin (BSA) at 20-25mg/ml in deionized water can be applied to the glass microscope slide (and coverslip) and allowed to evaporate off (to inhibit particle sticking to the glass surfaces).
  • For non-biological particles, 1% polyethylene glycol can be added to the microsphere solution as an anti-stick agent.
  • For biological particles the BSA application and Phosphate-Buffered Saline (PBS) (Sigma-Aldrich P5368-10PAK) can be used as anti-stick agents.
  • If particles begin to stick to the slide surfaces, a 3 minute sonication can increase the mobility of particles.

PBS Wikipedia

Students: If this recipe is inadequate (too many or too few particles) please revise.