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spinning_disk_confocal

Spinning Disk Confocal

Optics

Objectives
  1. Plan Fluor 10x /0.3 (Ph1 DL) 910 nm/pixel
  2. Plan Apo 40x/0.95 Corr (DIC N2 / 40X I) 227.5 nm/pixel
  3. Plan Apo VC 60x/1.2 WI
  4. Plan Apo VC 60x/1.4 Oil
  5. Plan Apo VC 100x/1.4 Oil (DIC N2 / 100X I) 91 nm/pixel
  6. Apo TIRF 100x/1.49 Oil 91 nm/pixel
Filter Turret 1 (FL1, Top)
  1. Empty
  2. DIC Analyzer
  3. GFP (long pass)
  4. YFP
  5. Rhodamine
  6. D/F/R
Yokogawa Dichroics
  1. 405/491/561/640
Emission Wheel

(To get NIS-Elements numbering, add 1 to these numbers)

  1. Open
  2. ET460/50m
  3. ET525/50m
  4. ET645/65m
  5. ET700/75m
  6. ET610/60m
  7. zET405/488/561/635m
  8. Empty
  9. Empty
  10. Empty

Hardware

  • Nikon Ti-E Microscope
  • Yokogawa CSU-22
  • Sutter Lambda LS-2 lamp for epifluorescence illumination
  • Sutter Emission wheel controlled by Lambda 10-B
  • Sutter TLED brightfield illumination
  • Photometrics Evolve Delta
  • Prior motorized stage with Piezo Z-drive
  • Okolab enclosure and Bold Line incubator

Lasers

  • 405 nm, 100 mW Coherent OBIS
  • 488 nm, 150 mW Coherent OBIS
  • 561 nm, 100 mW Coherent OBIS
  • 640 nm, 100 mW Coherent OBIS

Interface Requirements

  • 1 serial port for Sutter Lambda 10-B (COM7)
  • 1 serial port for Yokogawa CSU-22 (COM5)
  • 1 serial port for Prior Proscan II (COM2)
  • Firewire for Evolve
  • USB for Ti
  • USB for laser controller (COM4; MM only)

Firmware

  • Ti: Combination K
  • TiControl: 4.32

PC

  • Windows 7 64-bit
  • Core i7 3.4GHz
  • 16 GB RAM
  • NVidia GTX550Ti
  • 240GB SSD
  • 2TB Hard drive (RAID 1)

Software

  • Nikon Elements 5.02 build 1266
  • Micro-Manager 2.0
  • Micro-Manager 1.4

Laser Settings and Powers

Optimal AOTF settings, 5/22/2013:

Channel Frequency Amplitude
491 70.475 478.20
561 59.091 608.80
640 50.300 525.00

Laser Powers

Laser Power to Output 10x objective (mW)

Laser scroll bar
405 100.2
491 45.9
561 24
640 19
Date 405 nm 491 nm 561 nm 640 nm Notes
1/7/14 0.199 0.334 0.872 0.483
2/5/14 0.232 0.273 0.876 0.485
3/6/14 0.156 0.192 0.760 0.450
4/3/14 0.250 0.197 0.805 0.438
5/2/14 0.237 0.110 0.545 0.419
6/12/14 0.224 0.261 0.460 0.429
7/17/14 0.213 0.232 0.429 0.414
8/4/14 0.186 1.138 1.030 0.402 Direct modulation of lasers
8/15/14 0.126 1.02 1.16 0.576
9/9/14 0.119 1.02 1.27 0.498 realigned 488
10/9/14 0.107 0.985 1.12 0.461 realigned 488
11/6/14 0.113 0.957 1.237 0.480 new mirror mount for 488 laser line
12/4/14 0.111 1.118 1.214 0.465
1/5/15 0.292 2.94 1.675 0.730 realigned 488
2/2/15 0.301 2.83 1.77 0.696 realigned 488
3/5/15 0.297 2.62 1.61 0.690
4/2/15 0.292 2.58 1.62 0.681
4/16/15 2.45 realigned 488, 405 laser died
5/12/15 N/A 2.52 1.51 0.654 realigned 488
5/21/15 0.82 2.67 1.71 1.35 installed new 405 and 640 lasers
6/10/15 0.597 2.07 1.45 1.21
7/8/15 0.384 1.52 1.20 1.08 realigned 640, 561, 488
8/4/15 0.306 1.20 1.04 0.95 realigned 405
8/31/15 1.05 3.26 2.04 1.54 installed new fiber, realigned all lasers
10/14/15 1.00 2.98 1.87 1.50
11/14/15 0.94 3.00 1.98 1.46 realigned 561, 488
12/2/15 0.97 3.10 1.93 1.40 realigned 405, 488, 561, 640
1/6/16 0.97 2.15 1.82 1.31
1/20/16 0.89 2.60 1.80 1.42
2/4/16 0.87 2.80 1.83 1.35 realigned 488
3/1/16 0.93 2.73 1.80 1.33 realigned 488
3/30/16 0.78 2.50 1.75 1.37
5/9/16 0.80 2.26 1.82 1.45
6/15/16 0.77 2.27 1.78 1.41
/var/www/html/dokuwiki/data/pages/spinning_disk_confocal.txt · Last modified: 2017/11/03 09:02 by dlarsen