Instrument

Instrument name:

ZeissInverted confocal microscope(LSM 880) 

Room number:

1022 Instrument introduction Zeiss lsm880 inverted confocal fluorescence microscope is mainly used for long-term open field and fluorescence imaging of cultured cells or tissues. The system is equipped with airyscan detector which is unique to Zeiss, which can improve the resolution and imaging speed at the same time. Airyscan has a set of 32 array GaAs p-pmt detectors, without traditional pinholes. The 32 micro units of the sensor are like 32 pinholes, each sensing the light from the sample. Through the algorithm, the azimuth signals measured by the 32 micro units are summed up again to form a correct and clear image. Because there is no pinhole, the light signal is no longer covered by the pinhole. Airyscan can accept more light from the sample. Airyscan receives 1.25 times of the light from the traditional pinhole (1.25 AU). Each of its 32 micro units is equivalent to 0.2 Au, so it can greatly improve the signal-to-noise ratio of the image, and improve the resolution and detail brightness of the image. At 488 nm, airyscan has 1.7 times the detection capacity of the traditional confocal microscope. The XY resolution can reach 140 nm, and the z-axis resolution can reach 400 nm. 

Scope of application:

The inverted microscope is suitable for observing the living cells in the culture dish, equipped with DIC imaging mode, which can image the transparent cells. The living cell incubation system can be matched with 35mm and 60mm culture dishes and standard slides, which can be used for long-term continuous imaging of living cells and high-resolution imaging of fixed samples. It is suitable for observing and recording brain tissue sections, fluorescent labeling of living cells, three-dimensional image reconstruction, qualitative, quantitative, timing and location distribution detection of cell biological substances and ions, and analyzing the physiological and biochemical changes in brain tissues, cells or cells。 

Key technical indicators: 

Laser:

405nm(30mW)、561nm(20mW)、633nm(5mW)、Multiline argon ion green laser458/488/514nm(25mW)。Fast AOTF, 8 channels, switching time between multiple channels 5 microseconds。 

Objective:

10x (Air, NA=0.45)

20x (Air, NA=0.8)

25x (Water,Glycerin,Silicon oil,NA=0.8, WD=0.57mm)

40x (Air, NA=0.6, WD=3.48mm)

40x (Oil,NA=1.3, WD=0.2mm)

63x (Oil, NA=1.4, WD=0.19mm)

Imaging mode: 

General transmission light field imaging, DIC imaging, fluorescence imaging (DAPI, GFP, Cy3, Cy5). The minimum detection range of fluorescence spectrum is 3nm. 

DAPI: EX 365, BS FT 395, EM BP 445/50 

GFP: EX BP 470/40, BS FT 495, EM BP 525/50

CY3: EX BP 545/25, BS FT 570, EM BP 605/70

CY5: EX BP 640/30, BS FT 660, EM BP 690/50

The list of the most suitable dyes:

Channels

Dyes

DAPI

EX 365, BS FT 395, EM BP 445/50

  BFP (EBFP)(380\440)

  DAPI(359/461)

  DyLight 405(398\420)

  Hoechst 34580(392\440)

  LysoSensor Blue(374\424)

  LysoTracker Blue(373\422)

  Marina Blue(363\461)

  sgBFP™(387\451)

  Vybrant DyeCycle Violet(369\437)

GFP

EX BP 470/40, BS FT 495, EM BP 525/50

  BCECF (pH 5.5)(481\518)

  CFP2(490\510)

  Cy2™(492\507)

  Dendra2 (Green)(491\507)

  DiO(475\500)

  ecliptic pHluorin pH5.5(473\507)

  Emerald(491\511)

  FITC (Fluorescein)(495\519)

  Fluo-4(494\516)

  Fluorescein-pH 8.0(489\517)

  Fluoro-Emerald(494\518)

  GFP (EGFP)(489\511)

  LysoSensor Green(448\502)

  MitoTracker™ Green(490\512)

  NBD-X (MeOH)(467\538)

  NeuroTrace 500/525 Green Fluorescent Nissl        Stain(495\524)

  PKH67(488\500)

  SYTO 13(448\506)

  SYTO 16(489\520)

  SYTO 9(483\500)

  TurboGFP(482\503)

  wtGFP (wild type GFP, non-UV excitation)(474\509)

  YO-PRO-1(491\506)

  YOYO-1(491\508)

CY3

EX BP 545/25, BS FT 570, EM BP 605/70

  5-ROX (carboxy-X-rhodamine)(578\604)

  Alexa Fluor® 546(556\572)

  Alexa Fluor® 568(579\603)

  Amplex UltraRed peroxidation product (pH 7.5)(568\581)

  AsRed 2(576\592)

  ATTO 550(553\576)

  ATTO 565(563\592)

  BOBO™-3(570\605)

  BODIPY TMR-X(544\573)

  BODIPY TR-X (MeOH)(544\573)

  Cy3.5™(578\591)

  DsRed(559\583)

  DsRed-Expres(556\584)

  tdTomato(556\582)

  Ethidium bromide(518\603)

  Ethidium homodimer(527\617)

  KFP-Red(575\599)

  LOLO-1(568\580)

  LysoTracker Red(573\592)

  Magnesium Orange(550\574)

  mApple(566\594)

  Merocyanine 540(559\579)

  MitoTracker™ Orange(551\575)

  mRFP(555\583)

  mRuby(558\605)

  mStrawberry(574\596)

  mTangerine(568\585)

  Nile red-phospolipid(553\637)

  pHrodo™, succinimidyl ester(560\587)

  Rhod-2(553\577)

  Rhodamine phalloidin(558\575)

  Rhodamine Red-X(573\591)

  R-phycoerythrin (PE)(565\576)

  SNARF-1 488nm (ph 6.0)(548\586)

  SNARF-1 514nm (ph 6.0)(549\587)

  SpectrumOrange(554\587)

  Tetramethylrhodamine dextran(554\582)

  TRITC (Tetramethylrhodamine)(552\578)

  TurboRFP(553\573)

CY5

EX BP 640/30, BS FT 660, EM BP 690/50

  Alexa Fluor® 647(653\668)

  APC(651\660)

  APC (Allophycocyanin)(651\660)

  ATTO 647(644\670)

  ATTO 647N(645\668)

  BODIPY 650/665-X(646\664)

  Cy5™(649\666)

  DiD (DiIC18(5)) - Lipophilic Tracer(648\670)

  DyLight 649(651\673)

  FluoSpheres Dark Red fluorescent        microspheres(656\683)

  HiLyte Fluor™ 647(649\673)

  LIVE-DEAD Fixable Far Red Dead Cell  Stain(653\668)

  MitoTracker™ Deep Red(641\662)

  Red 650(649\665)

  SpectrumFRed (Far Red)(650\673)

  SYTO 60(650\681)

  TOTO-3(642\661)




 



Detector:

Built in two GaAsP ultra-high sensitivity linear detectors, two PMT linear detectors, high sensitivity monochromatic refrigeration CCD, one of which is the airyscan scanning mode detector. 

Resolving power:

In ultra-high resolution mode, the imaging speed is 5 frames / second (512 × 512 pixels, 16 bits) at 120nm in XY direction and 350nm in Z direction. Ultra high resolution fast imaging mode, 145nm in X direction, 180nm in Y direction, 450nm in Z direction, imaging speed 19 frames / second (512 × 512 pixels, 16 bits)。 

Scanning resolution:

The resolution can be freely selected from 4 × 1 to 8192 × 8192, and each channel can achieve the resolution of 8192 × 8192 and 16 bit gray level。 

Scanning speed: 

Galvanometer scanning galvanometer: 13 frames / second (512 × 512 pixels, 16 bits); 430 frames / second (512 × 16 pixels, 16 bits) 

Line scanning:

4-line / s to 6875 line / S (512 × 1 pixels, 16 bits) Fast scan mode: 27 frames / second (488 × 488 pixels, 16 bits) 

CCD acquisition:

The acquisition speed of full frame is 38 frames / S (1936 × 1460); spectrum monitoring range:400nm~1000nm; 

Sampling depth:

High quality drawing 14bit In airyscan scanning mode, there are high-resolution and high-speed scanning modes available: in high-resolution mode, the scanning speed is 7fps @ 512 × 512, and the scanning field is 22mm. Scanning speed 28fps @ 512 × 512 in high speed scanning mode。 

Scanning zoom:

The minimum optical zoom is 0.6x ~ 40x, which can be used with high-power objective lens for large field, high-speed and high-resolution scanning. 

Living cell incubation system:

Temperature control range: room temperature to 60 ℃, temperature regulation accuracy of 0.01 ℃ CO2 automatic mixing: concentration control range 0 to 8%, regulation accuracy 0.01% 

Humidification device:

200mlpureLarge capacity water humidification device, which can also be heated Incubation chamber: suitable for 35mm and 60mm culture dishes, standard slide, small eight hole culture tank 

Image processing software:

It has the function of spectrum scanning and resolution, which can remove the spontaneous fluorescence and fluorescence crosstalk; it has the function of free big picture splicing, CO location analysis and some basic measurement functions; it is equipped with the physiological analysis module, which has the graphical analysis of the average value of the fluorescence intensity of the region of interest, which can display and calculate the ion concentration in real time or after the completion of scanning; it has the histogram analysis tool, which can be measured It can measure the length, angle, area, fluorescence intensity and so on.


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