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='''Microbead loading'''= ==Procedures== *Start from the diluted and immobilized bead (with FITC labeled probes). Concentration: 1 mg/200 uL. ** The original bead info (Invitrogen): [[Media:Dynabead M-280.pdf|Dynabead M-280]]; [[Media:Dynabead application.pdf|Dynabead application]] ** 10 mg (6-7 x 10^8 beads) -> 1 mg = 6-7 x 10^7 beads *Resuspend by repeat pipetting using p200 pipettor. *Dilute in 1/200 ratio with nuclease-free H2O ** Take 1 uL resuspension into a fresh 1.5 mL tube with 199 uL Nuclease-free H2O. Mix gently by repeat pipetting (p200). ** The new bead conc. will be 6 x 10^7 bead / 200 (uL) x 1/200 = 1500 bead /uL *Leave the slide on Magnetic board. Transfer 10 uL, 20 uL, 40 uL on microwell arrays. ** The bead drop can barely cover the entire array area except using 20 ~ 40 uL of total volume. *Air day the slides in fume hood until all of the diluent are fully evaporated (More than 10 min will be needed to dry up) ** It may be possible to dry on on magnetic hot plate at 60 degree. ==Microscope images== *The structure microwell and microbead were characterized using Zessis A1-mode fluorescent microscope with 10X objective '''FIG. 1: Black and white image (10X objective)''' [[Image:blackwhite 2-13-09_bmp.bmp|600px|none]] '''FIG. 2: Color image in green (10X objective)''' [[Image:color 2-13-09_bmp.bmp|600px|none]] ==Discussion== *No bead was found on microwell arrays in both B&W and color images. It is possible that: # beads floated around on magnetic hot plate. The hot plate can't attract beads with enough magnetic force. # The dilution of bead is not optimized ideally. *Still some dust could be found on microwell arrays. # Higher conc. of HCL or longer soaking time could be applied in more tests in the furture.
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