MICROSCOPE MODE
SETUP & BRIGHTFIELD
FLUORESCENCE
PHASE CONTRAST
DIC
POLARISED LIGHT
DARK FIELD
CONFOCAL
SUPER-RES – STED
SUPER-RES – SMLM
INSERT
PHASE
DIC FILTER
POLARISING FILTER
DARKFIELD RING
HALOGEN LAMP
MERCURY LAMP
FOCUS
FIELD DIAPHRAGM
RIGHT EYEPIECE
OBJECTIVES
MOVE SLIDE
LIGHT INTENSITY
CONDENSER POSITIONING
CONDENSER FOCUS
CONDENSER APERTURE
LIGHT INTENSITY
MICROSCOPE MODE
SETUP & BRIGHTFIELD
FLUORESCENCE
PHASE CONTRAST
DIC
POLARISED LIGHT
DARK FIELD
CONFOCAL
SUPER-RES – STED
SUPER-RES – SMLM
CHANNEL 2
Ex 488 Em 500-550
100%
80V
60
0%
20V
-60
LASER
GAIN (HV)
OFFSET
CHANNEL 3
Ex 561 Em 570-600
100%
80V
60
0%
20V
-60
LASER
GAIN (HV)
OFFSET
CHANNEL 4
Ex 640 Em 650-700
100%
80V
60
0%
20V
-60
LASER
GAIN (HV)
OFFSET
Z-STACK & PINHOLE
TOP Z-POS
BOTTOM
Z-POS
no set
no set
RANGE
OPTICAL SECTION THICKNESS
LIVE VIEW
CAPTURE IMAGE
CAPTURE Z-STACK
SCAN RATE (FRAMES/SEC)
0.5
ZOOM
MICROSCOPE MODE
SETUP & BRIGHTFIELD
FLUORESCENCE
PHASE CONTRAST
DIC
POLARISED LIGHT
DARK FIELD
CONFOCAL
SUPER-RES – STED
SUPER-RES – SMLM
CHANNEL 1
Ex 633 Em 690-720
LASER
STED LASER
775 nm
LASER
AUTOSCALE
DECONVOLUTION
LIVE VIEW
CAPTURE IMAGE
PIXEL SIZE
120 nm
ZOOM
1x
6x
MICROSCOPE MODE
SETUP & BRIGHTFIELD
FLUORESCENCE
PHASE CONTRAST
DIC
POLARISED LIGHT
DARK FIELD
CONFOCAL
SUPER-RES – STED
SUPER-RES – SMLM
CHANNEL 1
Ex 647 Em 665-715
LASER
1000 µm
FOCUS
CAMERA SETTINGS
DATA ACQUISITION
ACTIVATION LASER
POWER
2%
DURATION
100
MS
EVERY
500
FRAMES
LIVE VIEW
CAPTURE IMAGE
CAPTURE SMLM IMAGE
DATA PROCESSING
INTENSITY THRESHOLD
LOW
HIGH
PROCESS SMLM IMAGE
DRIFT CORRECTION
EXPOSURE TIME
0ms
8ms
ACTIVITIES MENU
VISUALISING YOUR SAMPLE
OBJECTIVE LENS
PINHOLE
PIXEL DWELL
PIXEL ARRAYS
MULTICHANNEL IMAGES
3D & Z-STACKS
Excitation–emission: example for viewing GFP
Note how much brighter and blurrier the open pinhole image is and how much extra light comes from out of focus planes when the pinhole isn’t blocking it. This is obvious based on number of oversaturated red pixels.
×
Compare it with the images that you have just collected. If you close the pinhole too much, light can’t reach the detector.
×At a Pixel Dwell Time of 12 µseconds (compared to a dwell time of 2.0 µseconds), the time to collect photons is increased by a factor of 6x. This means the image will be much BRIGHTER and the image scan rate will be much SLOWER. Most common dwell times are around the 2–5 µsecond range.
×The resulting image is a merge of both AlexaFluor568 and AlexaFluor647.
Save this
image to compare it to a sequentially acquired image.
The resulting image is a merge of both AlexaFluor568 and AlexaFluor647 but scanned sequentially.
×Simultaneous
Sequential
×Note that each image is acquired frame by frame to ensure the absence of the spectral bleedthrough of the 568 into Channel 4 that was present in the simultaneous image.
×CONFOCAL
STED
×Use the zoom buttons to have a closer look into a specific region and drag over the image to pan.
When ready close this window to continue.
Lower (6%) STED laser intensity
Higher (35%) STED laser intensity
Higher (35%) STED laser intensity
(with autoscale)
Use the zoom buttons to have a closer look into a specific region and drag over the image to pan.
When ready close this window to continue.
Channel 3 3D image projection and rotation.
×Please wait while the image scans
Diagram
×
Important Points:
×Loading resources
For an optimal experience:
×If the simulator extends beyond the lower edge of your screen, drag the side edge of your browser until the simulator fits properly within the window.
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