Wiki source code of Confocal microscopy

Last modified by Jussi Kenkkilä on 2024/03/13 11:01
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1 (% id="HEquipment" class="section-title" %)
2 = Equipment =
3
4 {{toc maxLevel="2" minLevel="2"/}}
5
6 == Zeiss LSM 780 ==
7
8 (% style="color:#333333" %)Motorized Zeiss Axio Observer.Z1 inverted microscope.
9
10 (% style="color:#333333" %)[[image:https://www.helsinki.fi/assets/drupal/media-image/LSM780_1920_1200.jpg||alt="LSM 780" height="31%" width="50%"]]
11
12 === **Technical Information and capabilities** ===
13
14 ==== Objectives ====
15
16 (% class="relative-table wrapped" style="width:56.0%" %)
17 |=(((
18 Magnification
19 )))|=(((
20 Objective type
21 )))|=(((
22 NA
23 )))|=(% colspan="1" %)(% colspan="1" %)
24 (((
25 Working distance (mm)
26 )))|=(% colspan="1" %)(% colspan="1" %)
27 (((
28 phase contrast
29 )))|=(((
30 Immersion
31 )))
32 |(((
33 10x
34 )))|(((
35 Plan-Apochromat
36 )))|(((
37 0.45
38 )))|(% colspan="1" %)(% colspan="1" %)
39 (((
40 2.1
41 )))|(% colspan="1" %)(% colspan="1" %)
42 (((
43 -
44 )))|(((
45 Dry
46 )))
47 |(((
48 20x
49 )))|(((
50 Plan-Apochromat
51 )))|(((
52 0.80
53 )))|(% colspan="1" %)(% colspan="1" %)
54 (((
55 0.55
56 )))|(% colspan="1" %)(% colspan="1" %)
57 (((
58 -
59 )))|(((
60 Dry
61 )))
62 |(% colspan="1" %)(% colspan="1" %)
63 (((
64 25X
65 )))|(% colspan="1" %)(% colspan="1" %)
66 (((
67 LD LCI Plan-
68 Apochromat
69 )))|(% colspan="1" %)(% colspan="1" %)
70 (((
71 0.8
72 )))|(% colspan="1" %)(% colspan="1" %)
73 (((
74 0.57
75 )))|(% colspan="1" %)(% colspan="1" %)
76 (((
77 -
78 )))|(% colspan="1" %)(% colspan="1" %)
79 (((
80 Water, Oil, Glyc
81 )))
82 |(((
83 40x
84 )))|(((
85 Plan-Neofluor
86 )))|(((
87 1.30
88 )))|(% colspan="1" %)(% colspan="1" %)
89 (((
90 0.20
91 )))|(% colspan="1" %)(% colspan="1" %)
92 (((
93 -
94 )))|(((
95 Oil
96 )))
97 |(% colspan="1" %)(% colspan="1" %)
98 (((
99 40X
100 )))|(% colspan="1" %)(% colspan="1" %)
101 (((
102 C-Apochromat (on request)
103 )))|(% colspan="1" %)(% colspan="1" %)
104 (((
105 1.2
106 )))|(% colspan="1" %)(% colspan="1" %)
107 (((
108 0.28
109 )))|(% colspan="1" %)(% colspan="1" %)
110 (((
111 -
112 )))|(% colspan="1" %)(% colspan="1" %)
113 (((
114 Water
115 )))
116 |(((
117 63x
118 )))|(((
119 Plan-Apochromat
120 )))|(((
121 1.40
122 )))|(% colspan="1" %)(% colspan="1" %)
123 (((
124 0.19
125 )))|(% colspan="1" %)(% colspan="1" %)
126 (((
127 -
128 )))|(((
129 Oil
130 )))
131
132 (% id="HLaserlines" class="auto-cursor-target" %)
133 ==== Laser lines ====
134
135 (% class="relative-table wrapped" style="width:33.4613%" %)
136 |=(((
137 Laser
138 )))|=(((
139 Wavelength (nm)
140 )))|=(((
141 Example of fluorochromes
142 )))
143 |(((
144 Diode
145 )))|(((
146 405
147 )))|(((
148 DAPI, Hoechst
149 )))
150 |(((
151 Argon
152 )))|(((
153 458
154 )))|(((
155 CFP
156 )))
157 |(((
158 Argon
159 )))|(((
160 488
161 )))|(((
162 FITC, Alexa Fluor 488, GFP, Cy2, SytoxGreen, Acr. Orange/DNA
163 )))
164 |(((
165 Argon
166 )))|(((
167 514
168 )))|(((
169 YFP, Cy3
170 )))
171 |(((
172 Diode
173 )))|(((
174 561
175 )))|(((
176 PI, TRIT/Rhodamine, Mitotracker Orange, Alexa Fluor 546, DsRed2, Alexa Fluor 594, Texas Red, HcRed1
177 )))
178 |(((
179 Helium-Neon
180 )))|(((
181 633
182 )))|(((
183 Alexa 633, Cy5, APC, Alexa Fluor660, Alexa Fluor 647
184 )))
185
186 ==== Filters ====
187
188 Fluorescence filters for VIS-mode
189
190 (% class="wrapped" %)
191 |=(((
192 Zeiss filter set 49
193 )))|=(((
194 Zeiss filter set 38
195 )))|=(((
196 Zeiss filter set 43
197 )))
198 |(((
199 DAPI/ Hoechst
200 )))|(((
201 FITC/Alexa Fluor 488/ GFP
202 )))|(((
203 TRITC/ Cy3/ Alexa Fluor 546
204 )))
205
206 Transmitted light in Bright-field and DIC (ask for the DIC prisms if you need them).
207
208 ==== **Detectors** ====
209
210 * 2 PMT detectors (One with extra cooling for increased sensitivity in infrared)
211 * GaAsP spectral detector
212 * Transmitted light detector (T-PMT)
213
214 ==== **Software and accessories for live cell and multi-well imaging** ====
215
216 * Zeis ZEN 2010 software. Physiology
217 * FCS for GaAsP
218 * ROI HDR
219 * Image Correlation Spectroscopy analysis packages for ZEN 2010
220 * Definite Focus hardware for auto-focus system
221 * Heating Insert P S1 (temperature control)
222 * Incubator PM S1 (Temperature and CO,,2,, control)
223 * 40x and 63x water immersion objectives are available upon request
224
225 ==== System Changes ====
226
227 * Computer main drive was upgraded to a new bigger SSD (6.3.2024)
228
229 ==== Keywords ====
230
231 (% class="relative-table wrapped" style="width:53.6478%" %)
232 |=(((
233 Inverted microscope
234 )))|=(((
235 Multi-well
236 )))|=(((
237 Confocal
238 )))|=(((
239 Incubator
240 )))|=(% colspan="1" %)(% colspan="1" %)
241 (((
242 Environmental chamber
243 )))|=(% colspan="1" %)(% colspan="1" %)
244 (((
245 (% style="color:#4d4d4d" %)Definite focus
246 )))|=(% colspan="1" %)(% colspan="1" %)
247 (((
248 Water immersion
249 )))
250
251 (% id="HZeissLSM880" class="section-title" %)
252 == Zeiss LSM 880 ==
253
254 (% style="color:#555555" %)Zeiss inverted confocal microscope with GaAsP detector, AiryScan (Fast) detector, PicoQuant FLIM detector and a full environmental chamber.
255
256 [[image:https://www.helsinki.fi/assets/drupal/media-image/LSM880_user_1920_1200.jpg||alt="LSM 880" height="31%" width="50%"]]
257
258 === **Technical Information and capabilities** ===
259
260 ==== Objectives ====
261
262 (% class="wrapped" %)
263 |=(((
264 Magnification
265 )))|=(((
266 Objective type
267 )))|=(((
268 NA
269 )))|=(% colspan="1" %)(% colspan="1" %)
270 (((
271 (% style="color:#4d4d4d" %)Working distance (mm)
272 )))|=(% colspan="1" %)(% colspan="1" %)
273 (((
274 (% style="color:#4d4d4d" %)phase contrast
275 )))|=(((
276 Immersion
277 )))
278 |(((
279 10x
280 )))|(((
281 Plan-Apochromat
282 )))|(((
283 0.45
284 )))|(% colspan="1" %)(% colspan="1" %)
285 (((
286 2
287 )))|(% colspan="1" %)(% colspan="1" %)
288 (((
289 -
290 )))|(((
291 Dry
292 )))
293 |(((
294 20x
295 )))|(((
296 Plan-Apochromat
297 )))|(((
298 0.80
299 )))|(% colspan="1" %)(% colspan="1" %)
300 (((
301 0.55
302 )))|(% colspan="1" %)(% colspan="1" %)
303 (((
304 -
305 )))|(((
306 Dry
307 )))
308 |(% colspan="1" %)(% colspan="1" %)
309 (((
310 40x
311 )))|(% colspan="1" %)(% colspan="1" %)
312 (((
313 C-Apochromat
314 )))|(% colspan="1" %)(% colspan="1" %)
315 (((
316 1.20
317 )))|(% colspan="1" %)(% colspan="1" %)
318 (((
319 0.28
320 )))|(% colspan="1" %)(% colspan="1" %)
321 (((
322 -
323 )))|(% colspan="1" %)(% colspan="1" %)
324 (((
325 Water
326 )))
327 |(((
328 40x
329 )))|(((
330 Plan-Apochromat
331 )))|(((
332 1.40
333 )))|(% colspan="1" %)(% colspan="1" %)
334 (((
335 0.13
336 )))|(% colspan="1" %)(% colspan="1" %)
337 (((
338 -
339 )))|(((
340 Oil
341 )))
342 |(((
343 63x
344 )))|(((
345 Plan-Apochromat
346 )))|(((
347 1.40
348 )))|(% colspan="1" %)(% colspan="1" %)
349 (((
350 0.19
351 )))|(% colspan="1" %)(% colspan="1" %)
352 (((
353 -
354 )))|(((
355 Oil
356 )))
357
358 ==== Laser lines ====
359
360 (% class="relative-table wrapped" style="width:39.1662%" %)
361 |=(((
362 Laser
363 )))|=(((
364 Wavelength (nm)
365 )))|=(((
366 Example of fluorochromes
367 )))
368 |(((
369 Diode
370 )))|(((
371 405
372 )))|(((
373 DAPI, Hoechst
374 )))
375 |(((
376 Argon
377 )))|(((
378 458
379 )))|(((
380 CFP
381 )))
382 |(((
383 Argon
384 )))|(((
385 488
386 )))|(((
387 FITC, (% style="color:#333333" %)Alexa Fluor 488, GFP, Cy2, SytoxGreen, Acr.Orange/DNA
388 )))
389 |(((
390 Argon
391 )))|(((
392 514
393 )))|(((
394 (% style="color:#333333" %)YFP, Cy3
395 )))
396 |(((
397 Diode
398 )))|(((
399 561
400 )))|(((
401 (% style="color:#333333" %)PI, TRITC/Rhodamine, Mitotracker Orange, Alexa Fluor 546, DsRed2, Alexa Fluor 594, Texas Red, HcRed1
402 )))
403 |(((
404 Helium-Neon
405 )))|(((
406 633
407 )))|(((
408 Alexa 633, Cy5, APC, Alexa 660, Alexa 647
409 )))
410
411 ==== AiryScan Filters ====
412
413 * Plate
414 * BP 420-480 + BP 495-550
415 * BP 420-480 + BP 495-620
416 * BP 420-480 + LP 605
417 * BP 465-505 + LP 525
418 * BP 495-550 +LP 570
419 * BP 570-620 + LP 645
420
421 **Main Beam Splitters**
422
423 * MBS 458
424 * MBS 458/514
425 * MBS 458/461
426 * MBS 488
427 * MBS 488/561
428 * MBS 488/561/633
429 * MBS T89/R20
430 * Plate
431
432 **Secondary Beam Splitters**
433
434 * Mirror
435 * SBS LP 490
436 * SBS LP 515
437 * SBS LP 545
438 * SBS LP 635
439 * SBS SP 545
440 * SBS SP 610
441 * Plate
442
443 **Fluorescence filters for VIS-mode**
444
445 (% class="wrapped" %)
446 |=(((
447 Zeiss filter set 49
448 )))|=(((
449 Zeiss filter set 38
450 )))|=(((
451 Zeiss filter set 43
452 )))|=(((
453 Zeiss filter set 50
454 )))
455 |(((
456 DAPI/ Hoechst
457 )))|(((
458 FITC/ Alexa Fluor 488/ GFP
459 )))|(((
460 TRITC/ Cy3/ Alexa Fluor 546
461 )))|(((
462 Cy5, Alexa Fluor 647
463 )))
464
465 Transmitted light in Bright-field and DIC (ask for the DIC prisms if you need them).
466
467 ==== **Detectors** ====
468
469 * 2 PMT detectors (One with extra cooling for increased sensitivity in infrared)
470 * 1 GaAsP detector
471 Airyscan detector
472 Transmitted light detector (T-PMT)
473
474 ==== **Software and accessories for live cell and multi-well imaging** ====
475
476 * Zeis ZEN 2
477 * Modules: Experiment designer, Tiles & Positions, FRAP and FRET
478 * Definite Focus hardware autofocus system
479 * Scanning stage for multipoint or mosaic imaging
480 * Large environmental chamber
481 * Incubator insert PM S1 (Temperature and CO,,2,, control): Observation area 32 x 30 mm (oval) [[https:~~/~~/www.pecon.biz/products/product/?pid=5271%7CHeating_Insert_P_S>>url:https://www.pecon.biz/products/product/?pid=5271%7CHeating_Insert_P_S||shape="rect"]]
482 * 40x and 63x water immersion objectives are available upon request
483
484 ===== Environmental Chamber Temperature Setting =====
485
486 The readout of the temperature probe differs slightly from the media temperature in the sample chamber. To heat the sample to +37 C, adjust the Incubator to:
487
488 * H Insert P +38.0 °C
489 * H Unit XL +38.2 °C
490
491 ==== **Test your own fluorophores on this optical system** ====
492
493 To do so, click on the link below, choose the right configuration and add your own fluorophore. You will see the efficiency of the system for that specific label.
494
495 [[https:~~/~~/www.fpbase.org/microscope/fGbZLzo4KpjDLnCZqK9m6G/>>url:https://www.fpbase.org/microscope/fGbZLzo4KpjDLnCZqK9m6G/||shape="rect"]]
496
497 (% id="HKeywords-1" class="auto-cursor-target" %)
498 ==== Keywords ====
499
500 (% class="relative-table wrapped" style="width:72.2424%" %)
501 |=(((
502 (% style="color:#555555" %)Inverted microscope
503 )))|=(% colspan="1" %)(% colspan="1" %)
504 (((
505 Multi-well
506 )))|=(((
507 Confocal/ multiphoton
508 )))|=(((
509 Super-resolution
510 )))|=(% colspan="1" %)(% colspan="1" %)
511 (((
512 (% style="color:#4d4d4d" %)Incubator
513 )))|=(% colspan="1" %)(% colspan="1" %)
514 (((
515 (% style="color:#555555" %)Large environmental chamber
516 )))|=(% colspan="1" %)(% colspan="1" %)
517 (((
518 (% style="color:#4d4d4d" %)Definite focus
519 )))|=(% colspan="1" %)(% colspan="1" %)
520 (((
521 Water immersion
522 )))
523
524 (% id="HZeissLSM7MP" class="section-title" %)
525 == Zeiss LSM 7 MP ==
526
527 (% style="color:#333333" %)Upright multiphoton microscope optimized for intravital imaging of anaesthetized small laboratory animals.
528 The system allows acute as well chronic in vivo imaging experiments on internal (e.g. brain, kidney, liver etc.) and external organs (e.g. skin and eye).
529 The system can also be used for imaging fixed thick tissue samples.
530
531 (% style="color:#333333" %)[[image:LSM7_2_1920_1200.jpg||height="31%" width="50%"]]
532
533 === (% style="color:#333333" %)**Technical Information and capabilities**(%%) ===
534
535 ==== (% style="color:#333333" %)Objectives(%%) ====
536
537 (% class="relative-table wrapped" style="width:47.5152%" %)
538 |=(((
539 Magnification
540 )))|=(((
541 Objective type
542 )))|=(((
543 NA
544 )))|=(((
545 Work distance (mm)
546 )))|=(((
547 Immersion
548 )))|=(((
549 Coverglass thickness (mm)
550 )))
551 |(((
552 20x
553 )))|(((
554 W Plan-Apochromat
555 )))|(((
556 1.0
557 )))|(((
558 1.8
559 )))|(((
560 Water
561 )))|(((
562 0
563 )))
564 |(((
565 32x
566 )))|(((
567 C-Achroplan
568 )))|(((
569 0.85
570 )))|(((
571 0.85
572 )))|(((
573 Water
574 )))|(((
575 0-0.17
576 )))
577
578 ==== Lasers ====
579
580 * Coherent Chameleon Vision II, tunable wavelength: 690-1010nm
581 * APE Chameleon Compact OPO (optical parametric oscillator), tunable wavelength 1050-1300nm
582
583 (% id="HEmissionfilters" class="otsakepieni" %)
584 ==== Emission filters ====
585
586 * (% style="letter-spacing:0.0px" %)Reflected PMT's: BP 500-550 (green), BP 565-610 (red)
587 * Transmitted PMT's: BP 500-550 (green), BP 565-610 (red)
588 * Transmitted light filters for Bright-field and by request DoDt contrast (inclined illumination)
589
590 (% id="HLightSources" class="otsakepieni" %)
591 ==== Light Sources ====
592
593 * Widefield fluorescence and reflected light
594 * HXP 120 C metal halide lamp (fluorescence)
595 * LED-light source F1 (general-purpose light source)
596
597 (% class="otsakepieni" %)
598 (% style="color:#4d4d4d; letter-spacing:0.0px" %)**Detectors**
599
600 * 2 reflected light non-descanned PMT detectors with the possibility for filters
601 * 1 optional reflected light non-descanned nosepiece GaAsP detector without filters
602 * 2 transmitted light non-descanned PMT detectors with emission filters available on request
603 * 1 transmitted light non-descanned PMT detector available on request
604
605 **Software and accessories for live cell and multi-well imaging**
606
607 * ZEN 2011 software
608 * LinLab software for Scientifica motorized stage
609 * Dark microscope enclosure
610 * Univentor gas anesthesia system with mask, induction boxes and scavenger
611 * PhysioSuite hear/breath is an oxygenation monitoring system for mice and rats
612 * Animal's heating pad at the microscope stage to keep the animal warm during the anaesthesia
613 * Scientifica adjustable motorized microscope stage with a variety of intravital animal fixation adapters
614 * LED-light source F1 with flexible light guides for manipulations under the microscope
615 * Prior motorized microscope stage with multi-point scanning option available on request
616 * Dark environmental chamber (incubator XL) for minimization of stray light available on request
617 * Heating unit for temperature control of environmental chamber available on request.
618
619 (% id="HKeywords-2" class="otsakepieni" %)
620 ==== Keywords ====
621
622 (% class="relative-table wrapped" style="width:85.2727%" %)
623 |=(((
624 Upright microscope
625 )))|=(% colspan="1" %)(% colspan="1" %)
626 (((
627 Multi-well
628 )))|=(((
629 Wide-field
630 )))|=(((
631 Confocal/ multiphoton
632 )))|=(% colspan="1" %)(% colspan="1" %)
633 (((
634 (% style="color:#333333" %)Intravital imaging
635 )))|=(% colspan="1" %)(% colspan="1" %)
636 (((
637 (% style="color:#333333" %)In vivo imaging
638 )))|=(% colspan="1" %)(% colspan="1" %)
639 (((
640 Animal's heating pad
641 )))|=(% colspan="1" %)(% colspan="1" %)
642 (((
643 Dark environmental chamber
644 )))|=(% colspan="1" %)(% colspan="1" %)
645 (((
646 gas anesthesia
647 )))
648
649 (% id="HLeicaSP8CARS" class="section-title" %)
650 == Leica SP8 CARS ==
651
652 (% style="color:#555555" %)Leica inverted multi-photon microscope with two HyD detectors, resonant scanner, tunable CARS laser and a full environmental chamber.
653
654 [[image:https://www.helsinki.fi/assets/drupal/media-image/SP8CARS2_1920_1200.jpg||alt="SP8 CARS" height="31%" width="50%"]]
655
656 === **Technical Information and capabilities** ===
657
658 ==== Objectives ====
659
660 (% class="relative-table wrapped" style="width:60.6667%" %)
661 |=(((
662 Magnification
663 )))|=(((
664 Objective type
665 )))|=(% colspan="1" %)(% colspan="1" %)
666 (((
667 NA
668 )))|=(((
669 Working distance ((% style="color:#333333" %)µm)
670 )))|=(% colspan="1" %)(% colspan="1" %)
671 (((
672 Immersion
673 )))|=(% colspan="1" %)(% colspan="1" %)
674 (((
675 (% style="color:#4d4d4d" %)Coverglass thickness (mm)
676 )))|=(((
677 XY res. (nm)
678 )))|=(((
679 Z res. (nm)
680 )))
681 |(((
682 10x
683 )))|(((
684 HC PL APO CS
685 )))|(% colspan="1" %)(% colspan="1" %)
686 (((
687 0.40
688 )))|(((
689 360
690 )))|(% colspan="1" %)(% colspan="1" %)
691 (((
692 Multi
693 )))|(% colspan="1" %)(% colspan="1" %)
694 (((
695 -
696 )))|(((
697 488
698 )))|(((
699 4093
700 )))
701 |(((
702 25x
703 )))|(((
704 HCX IR APO L
705 )))|(% colspan="1" %)(% colspan="1" %)
706 (((
707 0.95
708 )))|(((
709 2400
710 )))|(% colspan="1" %)(% colspan="1" %)
711 (((
712 Water
713 )))|(% colspan="1" %)(% colspan="1" %)
714 (((
715 0.17
716 )))|(((
717 205
718 )))|(((
719 550
720 )))
721 |(((
722 20x
723 )))|(((
724 HC PL APO CS2
725 )))|(% colspan="1" %)(% colspan="1" %)
726 (((
727 0.75
728 )))|(((
729 680
730 )))|(% colspan="1" %)(% colspan="1" %)
731 (((
732 Multi
733 )))|(% colspan="1" %)(% colspan="1" %)
734 (((
735 -
736 )))|(((
737 260
738 )))|(((
739 1108
740 )))
741 |(((
742 40x
743 )))|(((
744 HC PL APO CS2
745 )))|(% colspan="1" %)(% colspan="1" %)
746 (((
747 1.10
748 )))|(((
749 650
750 )))|(% colspan="1" %)(% colspan="1" %)
751 (((
752 Water
753 )))|(% colspan="1" %)(% colspan="1" %)
754 (((
755 0.14-0.18
756 )))|(((
757 177
758 )))|(((
759 377
760 )))
761 |(((
762 63x
763 )))|(((
764 HC PL APO CS2
765 )))|(% colspan="1" %)(% colspan="1" %)
766 (((
767 1.20
768 )))|(((
769 300
770 )))|(% colspan="1" %)(% colspan="1" %)
771 (((
772 Water
773 )))|(% colspan="1" %)(% colspan="1" %)
774 (((
775 0.14-0.18
776 )))|(((
777 163
778 )))|(((
779 290
780 )))
781 |(((
782 63x
783 )))|(((
784 HC PL APO CS2
785 )))|(% colspan="1" %)(% colspan="1" %)
786 (((
787 1.30
788 )))|(((
789 300
790 )))|(% colspan="1" %)(% colspan="1" %)
791 (((
792 Glycerol
793 )))|(% colspan="1" %)(% colspan="1" %)
794 (((
795 0.14-0.19
796 )))|(((
797 150
798 )))|(((
799 276
800 )))
801
802 ==== Single-photon laser lines ====
803
804 (% class="relative-table wrapped" style="width:39.3308%" %)
805 |=(((
806 Laser
807 )))|=(((
808 Wavelength (nm)
809 )))|=(((
810 Example of fluorochromes
811 )))
812 |(((
813 (% style="color:#333333" %)Diode (Violet)
814 )))|(((
815 405
816 )))|(((
817 (% style="color:#333333" %)DAPI, Hoechst
818 )))
819 |(((
820 (% style="color:#333333" %)Argon (Blue)
821 )))|(((
822 458
823 )))|(((
824 (% style="color:#333333" %)CFP
825 )))
826 |(((
827 (% style="color:#333333" %)Argon (Blue)
828 )))|(((
829 476
830 )))|(((
831 -
832 )))
833 |(((
834 (% style="color:#333333" %)Argon (Blue)
835 )))|(((
836 488
837 )))|(((
838 (% style="color:#333333" %)FITC, Alexa Fluor 488, GFP, Cy2, SytoxGreen, Acr.Orange/DNA
839 )))
840 |(((
841 (% style="color:#333333" %)Argon (Blue)
842 )))|(((
843 496
844 )))|(((
845 -
846 )))
847 |(((
848 (% style="color:#333333" %)Argon (Blue)
849 )))|(((
850 514
851 )))|(((
852 (% style="color:#333333" %)YFP, Cy3
853 )))
854 |(((
855 (% style="color:#333333" %)DPSS (Lime)
856 )))|(((
857 561
858 )))|(((
859 (% style="color:#333333" %)PI, TRITC/Rhodamine, Mitotracker Orange, Alexa Fluor 546, DsRed2, Alexa Fluor 594, Texas Red, HcRed1
860 )))
861 |(((
862 (% style="color:#333333" %)Helium-Neon (Orange)
863 )))|(((
864 594
865 )))|(((
866 -
867 )))
868 |(% colspan="1" %)(% colspan="1" %)
869 (((
870 (% style="color:#333333" %)Helium-Neon (Red)
871 )))|(% colspan="1" %)(% colspan="1" %)
872 (((
873 633
874 )))|(% colspan="1" %)(% colspan="1" %)
875 (((
876 (% style="color:#333333" %)Alexa 633, Cy5, APC, Alexa 660, Alexa 647
877 )))
878
879 **Multiphoton laser (CARS, TPEF, SHG)**
880
881 * APE picoEmerald
882 * Wavelengths: 1064.5 nm and tunable 780-940 nm
883
884 ==== **Detectors** ====
885
886 * 2 high sensitivity hybrid detectors (HyD)
887 * 1 PMT detector
888 * 2 reflected light non-descanned PMT detectors with the possibility for filters
889 * 2 transmitted light non-descanned PMT detectors with the possibility for filters
890
891 ==== **Software and accessories for live cell and multi-well imaging** ====
892
893 * Full-enclosure type temperature-controlled environmental chamber for live-cell imaging and bleaching FRAP applications.
894 * An inverted temperature-controlled slow-flow imaging chamber available on request
895
896 ==== Keywords ====
897
898 (% class="wrapped" %)
899 |=(((
900 (% style="color:#555555" %)Inverted microscope
901 )))|=(% colspan="1" %)(% colspan="1" %)
902 (((
903 Multi-well
904 )))|=(((
905 CARS
906 )))|=(((
907 Confocal/(% style="color:#555555" %)multi-photon
908 )))|=(((
909 Live cell imaging
910 )))|=(((
911 Environmental chamber
912 )))|=(((
913 Water immersion
914 )))
915
916 (% id="HLeicaTCSSP8X" class="section-title" %)
917 == Leica TCS SP8 X ==
918
919 Leica TCS SP8 X with white light laser. Laser scanning confocal microscope for both fixed and live-cell imaging. Mounted on a motorized Leica inverted microscope.
920
921 [[image:https://www.helsinki.fi/assets/drupal/s3fs-public/migrated-generic-group-long-pages/body-images/78371-sp8x.jpg||alt="SP8 X" height="37%" width="50%"]]
922
923 === **Technical Information and capabilities** ===
924
925 ==== Objectives ====
926
927 (% class="relative-table wrapped" style="width:51.4545%" %)
928 |=(((
929 Magnification
930 )))|=(((
931 Objective type
932 )))|=(((
933 NA
934 )))|=(% colspan="1" %)(% colspan="1" %)
935 (((
936 (% style="color:#4d4d4d" %)Working distance (% style="color:#333333" %)(µm)
937 )))|=(((
938 Immersion
939 )))|=(((
940 Coverglass thickness (% style="color:#333333" %)(mm)
941 )))|=(((
942 XY res. (nm)
943 )))|=(((
944 Z res. (nm)
945 )))
946 |(((
947 10x
948 )))|(((
949 (% style="color:#333333" %)HC PL APO CS2
950 )))|(((
951 0.40
952 )))|(% colspan="1" %)(% colspan="1" %)
953 (((
954 2740
955 )))|(((
956 Dry
957 )))|(((
958 0.17
959 )))|(((
960 488
961 )))|(((
962 2630
963 )))
964 |(((
965 20x
966 )))|(((
967 (% style="color:#333333" %)HC PL APO CS2
968 )))|(((
969 0.75
970 )))|(% colspan="1" %)(% colspan="1" %)
971 (((
972 620
973 )))|(((
974 Dry
975 )))|(((
976 0.17
977 )))|(((
978 260
979 )))|(((
980 649
981 )))
982 |(((
983 40x
984 )))|(((
985 (% style="color:#333333" %)HC PL APO CS2
986 )))|(((
987 1.30
988 )))|(% colspan="1" %)(% colspan="1" %)
989 (((
990 240
991 )))|(((
992 Oil
993 )))|(((
994 0.17
995 )))|(((
996 150
997 )))|(((
998 299
999 )))
1000 |(((
1001 63x
1002 )))|(((
1003 (% style="color:#333333" %)HC PL APO CS2
1004 )))|(((
1005 1.40
1006 )))|(% colspan="1" %)(% colspan="1" %)
1007 (((
1008 140
1009 )))|(((
1010 Oil
1011 )))|(((
1012 0.17
1013 )))|(((
1014 139
1015 )))|(((
1016 236
1017 )))
1018 |(((
1019 63x
1020 )))|(((
1021 (% style="color:#333333" %)HC PL APO CS2
1022 )))|(((
1023 1.20
1024 )))|(% colspan="1" %)(% colspan="1" %)
1025 (((
1026 300
1027 )))|(((
1028 Water
1029 )))|(((
1030 0.14-0.18
1031 )))|(((
1032 163
1033 )))|(((
1034 290
1035 )))
1036
1037 ==== Lasers ====
1038
1039 * 405 nm diode
1040 * Supercontinuum laser, freely tunable between 470-670 nm, maximum of 8 simultaneous laser lines
1041
1042 ==== **Detectors, Software and accessories for live cell and multi-well imaging** ====
1043
1044 * 2 high sensitivity hybrid detectors (HyD)
1045 * 2 PMT detectors
1046 * 1 transmitted light detector (T-PMT)
1047
1048 The SP8 X has a full-enclosure type temperature-controlled environmental chamber for imaging live cells. While the chamber and microscope are heating up or cooling down the light path distances are constantly changing. Because of this it is not recommended to image while the chamber temperature is changing. To keep the chamber at a steady temperature, it should be either heated (+37 °C) or at room temperature for the whole day. The first reservation created for a given day determines the temperature of the system.
1049
1050 ==== Keywords ====
1051
1052 (% class="relative-table wrapped" style="width:49.8182%" %)
1053 |=(((
1054 Inverted microscope
1055 )))|=(((
1056 Multi-well
1057 )))|=(((
1058 Confocal
1059 )))|=(((
1060 (% style="color:#4d4d4d" %)Live cell imaging
1061 )))|=(% colspan="1" %)(% colspan="1" %)
1062 (((
1063 (% style="color:#4d4d4d" %)Environmental chamber
1064 )))|=(% colspan="1" %)(% colspan="1" %)
1065 (((
1066 White light laser
1067 )))
1068
1069 (% id="HLeicaStellaris8FALCON2FDLS" class="auto-cursor-target" %)
1070 == Leica Stellaris 8 FALCON / DLS ==
1071
1072 [[image:Leica_Stellaris_8_FALCON_1920_1200.jpg||alt="Stellaris FALCON" height="31%" width="50%"]]
1073
1074 Leica Stellaris 8 inverted confocal microscope with 5 sensitive HyD detectors and tunable white-light laser excitation source.
1075
1076 === Highlights: ===
1077
1078 * [[FALCON>>url:https://www.nature.com/articles/d42473-019-00261-x||shape="rect"]] module for fast fluorescence lifetime measurement
1079 * Digital Light-Sheet module
1080 * [[TauSense>>url:https://www.nature.com/articles/d42473-020-00364-w||shape="rect"]] detection
1081 * Fast resonant scanner
1082 * LASX Navigator
1083 * Leica [[Lightning>>url:https://www.leica-microsystems.com/science-lab/obtain-maximum-information-from-your-specimen-with-lightning/||shape="rect"]]
1084 * NIR excitation (up to 790nm) and detection (up to 850nm)
1085
1086 The [[LMU>>url:https://www2.helsinki.fi/en/infrastructures/bioimaging/light-microscopy-unit||shape="rect"]] have made a nice video covering the capabilities of the Stellaris 8 and its FALCON module (not DLS). Definitely worth a look:
1087
1088 === Technical information: ===
1089
1090 ==== Microscope ====
1091
1092 * Leica DMi8 inverted microscope
1093 * Motorized scanning stage
1094 * AFC harware autofocus
1095 * OKOlab full-enclosure incubator with temperature, CO,,2,,, and humidity control
1096
1097 ==== Objectives ====
1098
1099 (% class="relative-table wrapped" style="width:49.7532%" %)
1100 |=(% style="text-align: center;" %)(% style="text-align: center;" %)
1101 (((
1102 Magnification
1103 )))|=(% style="text-align: center;" %)(% style="text-align: center;" %)
1104 (((
1105 Objective type
1106 )))|=(% style="text-align: center;" %)(% style="text-align: center;" %)
1107 (((
1108 NA
1109 )))|=(% colspan="1" style="text-align: center;" %)(% colspan="1" style="text-align: center;" %)
1110 (((
1111 (% style="color:#4d4d4d" %)Working distance (% style="color:#333333" %)(µm)
1112 )))|=(% style="text-align: center;" %)(% style="text-align: center;" %)
1113 (((
1114 Immersion
1115 )))|=(% style="text-align: center;" %)(% style="text-align: center;" %)
1116 (((
1117 Coverglass thickness (% style="color:#333333" %)(mm)
1118 )))
1119 |(% style="text-align:center" %)(% style="text-align: center;" %)
1120 (((
1121 10x
1122 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1123 (((
1124 (% style="color:#333333" %)HC PL APO CS2
1125 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1126 (((
1127 0.40
1128 )))|(% colspan="1" style="text-align:center" %)(% colspan="1" style="text-align: center;" %)
1129 (((
1130 2740
1131 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1132 (((
1133 Dry
1134 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1135 (((
1136 0.17
1137 )))
1138 |(% style="text-align:center" %)(% style="text-align: center;" %)
1139 (((
1140 20x
1141 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1142 (((
1143 (% style="color:#333333" %)HC PL APO CS2
1144 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1145 (((
1146 0.75
1147 )))|(% colspan="1" style="text-align:center" %)(% colspan="1" style="text-align: center;" %)
1148 (((
1149 620
1150 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1151 (((
1152 Dry
1153 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1154 (((
1155 0.17
1156 )))
1157 |(% style="text-align:center" %)(% style="text-align: center;" %)
1158 (((
1159 40x
1160 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1161 (((
1162 (% style="color:#333333" %)HC PL APO CS2 CORR
1163 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1164 (((
1165 1.25
1166 )))|(% colspan="1" style="text-align:center" %)(% colspan="1" style="text-align: center;" %)
1167 (((
1168 350
1169 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1170 (((
1171 Glycerol
1172 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1173 (((
1174 0.14-0.18
1175 )))
1176 |(% style="text-align:center" %)(% style="text-align: center;" %)
1177 (((
1178 63x
1179 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1180 (((
1181 (% style="color:#333333" %)HC PL APO CS2
1182 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1183 (((
1184 1.40
1185 )))|(% colspan="1" style="text-align:center" %)(% colspan="1" style="text-align: center;" %)
1186 (((
1187 140
1188 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1189 (((
1190 Oil
1191 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1192 (((
1193 0.17
1194 )))
1195 |(% style="text-align:center" %)(% style="text-align: center;" %)
1196 (((
1197 63x
1198 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1199 (((
1200 (% style="color:#333333" %)HC PL APO CS2 motCORR
1201 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1202 (((
1203 1.20
1204 )))|(% colspan="1" style="text-align:center" %)(% colspan="1" style="text-align: center;" %)
1205 (((
1206 300
1207 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1208 (((
1209 Water
1210 )))|(% style="text-align:center" %)(% style="text-align: center;" %)
1211 (((
1212 0.14-0.18
1213 )))
1214
1215 ==== Lasers ====
1216
1217 * 405nm diode laser
1218 * supercontinuum laser, freely tunable between 440-790nm, maximum of 8 simultaneous laser lines
1219
1220 ==== Scanner ====
1221
1222 Tandem scanner with both traditional point scanner and fast 8kHz resonant scanner.
1223
1224 Five sensitive HyD detectors:
1225
1226 * 2 HyD S - all round detector for high performance detection throughout the spectrum
1227 * 2 HyD X - specialized for fast fluorescence lifetime imaging in the visible to the far-red
1228 * 1 HyD R - specialized for detection above 740nm (incl. fast fluorescence lifetime)
1229
1230 Please note that all 5 detectors are suitable for regular confocal imaging. However, the **HyD X**'s and **HyD R** are **not** tolerant of over exposure. If overexposed they will shut down and may be **permanently damaged**. You can find out more about the Leica HyD detectors and their capabilities [[here>>url:https://www.nature.com/articles/d42473-020-00398-0||shape="rect"]].
1231
1232 ==== Software Leica LAS X ====
1233
1234 * --4.5.0--
1235 * 4.6.0 (5.10.2023)
1236
1237 == (% style="color:#333333" %)Andor Dragonfly(%%) ==
1238
1239 (% style="color:#333333" %)[[image:https://www.helsinki.fi/assets/drupal/media-image/Andor_Dragonfly_1920_1200.jpg||alt="Andor Dragonfly" height="31%" width="50%"]]
1240
1241 === Technical Information and capabilities ===
1242
1243 Andor [[Dragonfly>>url:https://andor.oxinst.com/products/dragonfly||shape="rect"]] 505
1244
1245 ==== Objectives ====
1246
1247 (% class="relative-table wrapped" %)
1248 |=(((
1249 Magnification
1250 )))|=(((
1251 Objective type
1252 )))|=(((
1253 NA
1254 )))|=(((
1255 Working distance (mm)
1256 )))|=(% colspan="1" %)(% colspan="1" %)
1257 (((
1258 (% style="color:#4d4d4d" %)Phase-contrast
1259 )))|=(% colspan="1" %)(% colspan="1" %)
1260 (((
1261 Immersion
1262 )))|=(% colspan="1" %)(% colspan="1" %)
1263 (((
1264 (% style="color:#4d4d4d" %)PFS
1265 )))
1266 |(((
1267 10x
1268 )))|(((
1269 Plan Fluor DL
1270 )))|(((
1271 0.30
1272 )))|(((
1273 15.2
1274 )))|(% colspan="1" %)(% colspan="1" %)
1275 (((
1276 Ph1
1277 )))|(% colspan="1" %)(% colspan="1" %)
1278 (((
1279 Dry
1280 )))|(% colspan="1" %)(% colspan="1" %)
1281 (((
1282 +
1283 )))
1284 |(((
1285 20x
1286 )))|(((
1287 Plan Apo λ
1288 )))|(((
1289 0.75
1290 )))|(((
1291 1.0
1292 )))|(% colspan="1" %)(% colspan="1" %)
1293 (((
1294 -
1295 )))|(% colspan="1" %)(% colspan="1" %)
1296 (((
1297 Dry
1298 )))|(% colspan="1" %)(% colspan="1" %)
1299 (((
1300 +
1301 )))
1302 |(((
1303 40x ^^1), 2)^^
1304 )))|(((
1305 Apo LWD
1306 )))|(((
1307 1.15
1308 )))|(((
1309 0.6
1310 )))|(% colspan="1" %)(% colspan="1" %)
1311 (((
1312 -
1313 )))|(% colspan="1" %)(% colspan="1" %)
1314 (((
1315 Water
1316 )))|(% colspan="1" %)(% colspan="1" %)
1317 (((
1318 +
1319 )))
1320 |(((
1321 60x
1322 )))|(((
1323 Plan Apo VC
1324 )))|(((
1325 1.2
1326 )))|(((
1327 0.29
1328 )))|(% colspan="1" %)(% colspan="1" %)
1329 (((
1330 -
1331 )))|(% colspan="1" %)(% colspan="1" %)
1332 (((
1333 Water
1334 )))|(% colspan="1" %)(% colspan="1" %)
1335 (((
1336 +
1337 )))
1338 |(((
1339 100x ^^1)^^
1340 )))|(((
1341 SR Apo TIRF
1342 )))|(((
1343 1.49
1344 )))|(((
1345 0.12
1346 )))|(% colspan="1" %)(% colspan="1" %)
1347 (((
1348 -
1349 )))|(% colspan="1" %)(% colspan="1" %)
1350 (((
1351 Oil
1352 )))|(% colspan="1" %)(% colspan="1" %)
1353 (((
1354 +
1355 )))
1356 |(% colspan="1" %)(% colspan="1" %)
1357 (((
1358 40x
1359 )))|(% colspan="1" %)(% colspan="1" %)
1360 (((
1361 Plan Apo λ
1362 )))|(% colspan="1" %)(% colspan="1" %)
1363 (((
1364 1.25
1365 )))|(% colspan="1" %)(% colspan="1" %)
1366 (((
1367 0.3
1368 )))|(% colspan="1" %)(% colspan="1" %)
1369 (((
1370 -
1371 )))|(% colspan="1" %)(% colspan="1" %)
1372 (((
1373 Silicone oil
1374 )))|(% colspan="1" %)(% colspan="1" %)
1375 (((
1376 +
1377 )))
1378
1379 ^^1)^^ Not Plan corrected
1380
1381 ^^2)^^ Suitable for NIR labels. Chromatic correction 405-950nm
1382
1383 ==== Lasers ====
1384
1385 * 404nm, 100mW
1386 * 488nm, 150mW
1387 * 561nm, 100mW
1388 * 642nm, 110mW
1389 * 730nm, 30mW
1390
1391 ==== Filters ====
1392
1393 ===== For visual inspection of your sample through the eyepiece =====
1394
1395 * DAPI-50606
1396 * GFP-30309
1397 * TRITC-B 05
1398 * mCherry-C
1399
1400 ===== For imaging =====
1401
1402 * 445 -46 nm
1403 * 521 -38 nm
1404 * 594 -43 nm
1405 * 685 -47nm
1406 * 809 -90nm
1407 * TR-POL-DIC-FWL, Polarizer
1408 * 445/521/594/685/809
1409
1410 ==== Cameras ====
1411
1412 * Andor Zyla 4.2 sCMOS
1413 * Andor iXon 888 U3 EMCCD
1414
1415 ==== Software ====
1416
1417 * Fusion 2.0
1418 ** [[ClearView-GPU>>url:https://andor.oxinst.com/learning/view/article/clearview-gpu-deconvolution-put-those-photons-back-where-they-came-from||shape="rect"]] online deconvolution
1419 ** iXon [[SRRF-Stream>>url:https://andor.oxinst.com/products/srrf-stream||shape="rect"]] real-time super-resolution
1420 * Bitplane Imaris
1421
1422 ==== **Test your own fluorophores on this optical system** ====
1423
1424 To do so, click on the link below, choose the right configuration and add your own fluorophore. You will see the efficiency of the system for that specific label.
1425
1426 [[https:~~/~~/www.fpbase.org/microscope/MHoNXQQFSXUuAowQ34PYg8/?d=andor-ixon-life-888-bv>>url:https://www.fpbase.org/microscope/MHoNXQQFSXUuAowQ34PYg8/?d=andor-ixon-life-888-bv||shape="rect"]]
1427
1428 ==== Data Transfers From the Dragonfly Computer ====
1429
1430 If you're exporting or transferring data from the Dragonfly computer and not using the microscope, please start the Fusion OFFLINE version from the desktop. Using the regular Fusion while the microscope is off will confuse it and cause problems to the next user.