Skip to main content

Table 1 Summary of imaging parameters (Zeiss LSM510/META/NLO specifications)

From: Three dimensional multiphoton imaging of fresh and whole mount developing mouse mammary glands

Method

Laser

Emission

Imaging parameters

Image

Excitation

Live Tissue

860 nm

390-465 nm

Ch2 BP 390–465 IR

Figure 2

(5.8%)

500-550 nm

Ch3 BP 500–550 IR

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

Live Tissue

860 nm

390-465 nm

ChS BP 393-436

Figure 3A-C

(5.8%)

 

ChD

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

 

860 nm

390-465 nm

ChS BP 393-436

Figure 3D

(5.8%)

500-550 nm

Ch2 BP 500-550

ChD

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

Live Tissue

860 nm

390-465 nm

Ch2 BP 390–465 IR

Figure 4 (Live Tissue)

(5.8%)

500-550 nm

Ch3 BP 500–550 IR

ChD

ChD (unfiltered)

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

Carmine Alum WM

860 nm

390-465 nm

Ch2 BP 390–465 IR

Figure 4 (Whole Mount)

(5.8%)

500-550 nm

Ch3 BP 500–550 IR

ChD

ChD (unfiltered)

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

Carmine Alum WM

543 nm (50.5%)

565-615 nm

Ch3 BP 565–615 IR

Figure 5A,B

ChD

MBS: HFT 488/543/633

DBS1: NFT 635 VIS

DBS2: NFT 545

FW1: None

Carmine Alum WM

750 nm (2.1%)

565-615 nm

Ch3 BP 565–615 IR

Figure 5C

MBS: HFT KP650

DBS1: NFT KP545

DBS2: NFT 545

FW1: None

BF NDD

Carmine Alum WM

750 nm (5.8%)

565-615 nm

Ch3: BP 565–615 IR

Figure 6

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 545

FW1: None

Carmine Alum WM

860 nm (5.8%)

390-465 nm

Ch2 BP 390–465 IR

Figure 7

650-710 nm

Ch3 BP 650–710 IR

ChD

ChD (unfiltered)

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

Carmine Alum WM

860 nm (5.8%)

393-436

ChS BP 393-436

Figure 7B

ChD

ChD (unfiltered)

SHG-B/SHG-F image only

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

Carmine Alum WM

860 nm (5.8%)

390-465 nm

Ch2 BP 390–465 IR

Figure 8

650-710 nm

Ch3 BP 650–710 IR

ChD

ChD (unfiltered)

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

Carmine Alum WM

860 nm (5.8%)

390-465 nm

Ch2 BP 390–465 IR

Figure 9

650-710 nm

Ch3 BP 650–710 IR

ChD

ChD (unfiltered)

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

Carmine Alum WM

750, 800, 830, 860, 890, 950 nm (0.1%)

Lambda scan

ChS:361-704

Figure 10

361-704 nm

MBS: HFT KP650

DBS1: None

DBS2: None

FW1: None

Carmine Alum WM

860 nm (5.8%)

390-465 nm

Ch2 BP 390–465 IR

Figure 11 (Ex 860/ Em 650–710)

650-710 nm

Ch3 BP 650–710 IR

ChD

ChD (unfiltered)

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

 

750 nm (5.8%)

565-615 nm

Ch3: BP 565–615 IR

Figure 11 (Ex 750/ Em 565–615)

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 545

FW1: None

Unstained WM

735, 860, 960 nm (0.1%)

Lambda scan

ChS:361-704

Figure 12A, B

361-704 nm

MBS: HFT KP650

DBS1: None

DBS2: None

FW1: None

 

860 nm (5.8%)

390-465 nm

Ch2 BP 390–465 IR

Figure 12C-E

500-530 nm

Ch3 BP 500–530 IR

or

or

650-710 nm ChD

Ch3 BP 650–710 IR ChD (unfiltered)

MBS: HFT KP650

 

DBS1: Mirror

DBS2: NFT 490

FW1: None

Unstained WM

860, 890 nm (5.8%)

390-465 nm

Ch2 BP 390–465 IR

Additional file 13: Figure S11

500-530 nm

Ch3 BP 500–530 IR

ChD

ChD (unfiltered)

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

 

800, 890 nm (0.1%)

Lambda scan

ChS:361-704

Additional file 13: Figure S11

361-704 nm

MBS: HFT KP650

DBS1: None

DBS2: None

FW1: None

Unstained WM

800, 890 nm (0.1%)

390-465 nm

Ch2 BP 390–465 IR

Additional file 13: Figure S11

500-530 nm

Ch3 BP 500–530 IR

ChD

ChD (unfiltered)

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

Carmine Alum WM

860 nm (5.8%)

390-465 nm

Ch2 BP 390–465 IR

Additional file 1: Figure S1

650-710 nm

Ch3 BP 650–710 IR

ChD

ChD (unfiltered)

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

Live Tissue

860 nm (0.1%)

Lambda scan

ChS:361-704

Additional file 2: Figure S2

361-704 nm

MBS: HFT KP650

DBS1: None

DBS2: None

FW1: None

Carmine Alum WM

860 nm (5.8%)

390-465 nm

Ch2 BP 390–465 IR

Additional file 4: Figure S3

650-710 nm

Ch3 BP 650–710 IR

ChD

ChD (unfiltered)

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

Carmine Alum WM

860, nm (5.8%)

390-465 nm

Ch2 BP 390–465 IR

Additional file 5: Figure S4

650-710 nm

Ch3 BP 650–710 IR

ChD

ChD (unfiltered)

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

Carmine Alum WM

860 nm

Lambda scan

ChS:361-704

Additional file 7: Figure S5A,B

(0.1%)

361-704 nm

MBS: HFT KP650

Additional file 11: Figure S9B

DBS1: None

DBS2: None

FW1: None

Carmine Alum WM

860 nm (5.8%)

390-465 nm

Ch2 BP 390–465 IR

Additional file 7: Figure S5C

650-710 nm

Ch3 BP 650–710 IR

or

or

565-615

Ch3 BP 565–615 IR

ChD

ChD (unfiltered)

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

Carmine Alum WM

860 nm (5.8%)

390-465 nm

Ch2 BP 390–465 IR

Additional file 8: Figure S6

650-710 nm

Ch3 BP 650–710 IR

or

or

565-615

Ch3 BP 565–615 IR

ChD

ChD (unfiltered)

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

Carmine Alum WM

860 nm (5.8%)

390-465 nm

Ch2 BP 390–465 IR

Additional file 9: Figure S7

650-710 nm

Ch3 BP 650–710 IR

or

or

565-615

Ch3 BP 565–615 IR

ChD

ChD (unfiltered)

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

Unstained WM

860 nm

Lambda scan

ChS:361-704

Additional file 10: Figure S8

(0.1%)

361-704 nm

MBS: HFT KP650

DBS1: None

DBS2: None

FW1: None

Unstained WM

860 nm (0.1%)

390-465 nm

Ch2 BP 390–465 IR

Additional file 11: Figure S9A, C

500-530 nm

Ch3 BP 500–530 IR

ChD

ChD (unfiltered)

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

Unstained WM

860 nm (0.1%)

390-465 nm

Ch2 BP 390–465 IR

Additional file 12: Figure S10

500-530 nm

Ch3 BP 500–530 IR

ChD

ChD (unfiltered)

MBS: HFT KP650

DBS1: Mirror

DBS2: NFT 490

FW1: None

  1. Filters and Dichroics (LSM 510 Manual).
  2. KP = Low Pass Filter. Example: KP 685 passes wavelengths less than 685 nm to the detector.
  3. LP = High Pass Filter. Example: LP 505 passes wavelengths higher than 505 nm to the detector.
  4. BP = Bandpass Filter. Example: BP 565–615 passes wavelengths 565 nm to 615 nm to the detector.
  5. HFT = Main Dichroic Beam Splitter. Example: HFT 488/543 Excitation wavelengths (488 and 543 nm) sent to the sample.
  6. Passes all other wavelengths up to the detection channels. Example: HFT KP 700/488 Works as both a dichroic and a low pass filter. Will pass all wavelengths less than 700 nm, except for 488 nm, which hits sample, but is not sent to detector.
  7. NFT = Secondary Dichroic Beam Splitter. Example: NFT 543 Wavelengths above 543 nm, pass straight through. Wavelengths less than this amount are reflected 90 degrees.
  8. NT = Neutral Density Filter. Example: Works as an attenuation filter. NT 80/20 passes 80% of the light, cuts 20% of light.
Back to article page

BMC Cancer

ISSN: 1471-2407

Contact us