HANYANG UNIVERSITYº¸À¯±â±â

º¸À¯±â±â¸®½ºÆ®

Ȩ  >  º¸À¯±â±â¸®½ºÆ®  >  º¸À¯±â±â¸®½ºÆ®
È­ÇÕ¹°Àüó¸®/ºÐ¼®Àåºñ
±¤ÇÐ/ÀüÀÚ¿µ»óÀåºñ
¹°¸®ÀûÃøÁ¤Àåºñ
ÃÊ°í¼Ó À¯¼¼Æ÷ ÀÚµ¿ºÐ¸®±â
ÃÊ°í¼Ó À¯¼¼Æ÷ ÀÚµ¿ºÐ¸®±â
±â±â ¼³¸í
Àåºñ¸í(¾à¾î) Automated High-speed Cytometry Sorter
¸ðµ¨¸í FACSAriaIII
ºê·£µå Becton Dickinson(BD)
À§Ä¡ ½Å¼ÒÀç°øÇаü 124È£
´ã´çÀÚ ±è¿¬È£ yonoli@hanyang.ac.kr
´ã´çÀÚ ÀüÈ­¹øÈ£ 02-2220-2757
±â±â»óÅ ¿¹¾à°¡´É
ÀμâÇϱâ¸ñ·ÏÀ¸·Î

¿ø¸®¿Í Ư¡

FACS(Fluorescence activated cell sorter)¶ó´Â ±â°è´Â Çü±¤¹°Áú·Î ¿°»öµÈ ¼¼Æ÷¸¦ ±â°è ÀåÄ¡¼Ó¿¡¼­ sheath ¿ë¾×°ú ÇÔ²² Èê·Á º¸³»¸é¼­ ƯÁ¤ ÆÄÀåÀÇ laser beamÀ» Á¶»çÇÏ¿©, °¢ ¼¼Æ÷ÀÇ Æ¯¼º(Å©±â, complexity, 7°¡Áö ÆÄÀåÀÇ Çü±¤¹ß±¤µµ)¸¦ ºÐ¼®ÇÒ ¼ö ÀÖ´Â ÀåÄ¡ÀÌ´Ù. ºÐ¼® ÈÄ Æ¯¼ºÀÌ °°Àº ¼¼Æ÷¸¸À» ºÐ¸®ÇØ ³¾ ¼ö ÀÖ´Â ÀåÄ¡µµ ÀÖ´Ù. ±×·¯¹Ç·Î FACS ±â°è´Â Å©°Ô ºÐ¼®¿ë(¿¹: FACS Calibur, FACS Canto)¿Í ¼¼Æ÷ºÐ¸®¿ë(¿¹: FACS Aria sorter)·Î ³ª´¶´Ù. º» ¿øÀÇ ±â±â´Â ¼¼Æ÷ºÐ¸® Àü¿ëÀÎ FACS Aria ¥² sorterÀÌ´Ù.

»ç¾ç

1. Optics

1) Excitation Optics

- Fixed optical alignment of all Class IIIb lasers upon the cuvette flow cel

- 488-nm and 633-nm lasers come standard All other laser choices are optional.

- All are solid state except the 633 nm which is gas.

- Beam height: 9 ¡¾3 ¥ìm

- Beam width: 65 ¡¾7 ¥ìm

2) Emission Optics

        2-1) Optical Coupling

- The quartz cuvette flow cell is gel-coupled by refractive index-matching optical gel to the fluorescent objective lens for

  optimal collection efficiency

- Numerical aperture: 1.2

 2-2) Forward Scatter Detector and Filters

           - Photodiode with 488/10 bandpass filter for the 488-nm laser and 462/52 bandpass filter for the 445-nm laser.

 2-3) Side Scatter Detector

           - Photomultipler with a 488/10 bandpass filter for the 488-nm laser and a 445/15 bandpass filter for the 445-nm laser

 2-4) Fluorescence Detectors and Filters

           - Five fixed-fiber apertures for the 488, 633, 561, 445, and 405-nm lasers

 

2. Fluidics

1) Nozzles

- 70, 85, 100¥ìm, removable and can be sonicated

2) Automatic Cell Deposition Unit

- ACDU for slide and plate sorting : 6, 24, 48, 96, and 384-well plates

 

3. Performance

1) Sort Performance

       3-1) Drop Drive Frequency

- Range : 1 - 100,000 Hz

       3-2) Purity and Yield

- At 70 psi and 90 kHz with an average threshold rate of 25,000 events per second, a four- way sort achieved a purity of >98% and a yield >80% of Poisson¡¯s expected yield

- Higher threshold rates up to 70,000 events per second can be achieved without affecting purity ; however, yield will decrease based on Poisson statistics

       3-3) Viability

- Sorts were performed at a variety of sheath pressures using cell lines and human peripheral

blood mononuclear cells (PBMCs)

- All sorts resulted in cells that proved viable and proliferated for several days post-sort

       3-4) Sort Collection Devices

          - Two-way sorting : 12 x 75 mm and 15 mL

          - Four-way sorting: 1.5 mL microtube and 12 x 75 mm

       3-5) Sort Collection Cooling

          - Water recirculator for refrigeration/heating

½Ã·áÁغñ¹æ¹ý


»ç¿ë·á

20,000¿ø/30ºÐ + 30,000¿ø(±âº»)