Confocal Microscopy

Confocal Microscopy

Confocal microscopy is a fluorescence optical imaging technique for increasing optical resolution and contrast of a micrograph. This technique uses a laser point source to scan the sample and a pinhole to reduce collection of light from outside the focal plane and allows investigators to acquire thin optical sections at various focal planes. Stacks of images can be acquired for 3D visualization.


Application examples:

  • Life Sciences: wide range of imaging applications including the study of the spatial distributions of macromolecules in either fixed or living cells (e.g., protein localization analysis), automated collection of 3D data, the imaging of multiple labeled specimens and the measurements of physiological events in living cells (e.g., cell cycle studies).  
  • Material Sciences: used for investigating polymers, coatings, emulsions, etc.

 

We operate with the Zeiss LSM 710, a confocal laser scanning microscope able to generate high-resolution 3D images of specimens with high sensitivity and low photodamage.


 

System components:

LASERs

Laser Unit

Wavelength

Maximum Power

Diode 405-30

405 nm

30 mW

Argon

458, 488 and 514 nm

25 mW

DPSS 561-10

561 nm

15 mW

HeNe633

633 nm

5 mW

 

Objectives

Magnification

Model

Immersion

10x

EC Plan-Neofluar            

air

40x

EC Plan-Neofluar

oil

63x

Plan-Apochromat

oil

 

Filter sets (ocular)

Reflectors

Position/name

Excitation

Dichroic

Emission

Green

1-38 GFP

450-490 nm

495 nm

500-550 nm

Red

2-43 DsRed

533-558 nm

570 nm

570-640 nm

Blue

3-49 DAPI

G 365 nm

395 nm

420-470 nm

 

Software

Image acquisition and analysis is performed using the Zen 2010 software that includes the following modules:

  • Z stack
  • Time Series
  • Bleaching
  • Tile Scan
  • Positions
  • Regions
Publications with our Zeiss LSM 710:

Araújo, D. et al. Chitin-Glucan Complex Hydrogels: Physical-Chemical Characterization, Stability, In Vitro Drug Permeation, and Biological Assessment in Primary Cells. Polymers (Basel). 15, (2023).
https://doi.org/10.3390/polym15040791


Confocal Microscopy

Confocal microscopy is a fluorescence optical imaging technique for increasing optical resolution and contrast of a micrograph. This technique uses a laser point source to scan the sample and a pinhole to reduce collection of light from outside the focal plane and allows investigators to acquire thin optical sections at various focal planes. Stacks of images can be acquired for 3D visualization.


Application examples:

  • Life Sciences: wide range of imaging applications including the study of the spatial distributions of macromolecules in either fixed or living cells (e.g., protein localization analysis), automated collection of 3D data, the imaging of multiple labeled specimens and the measurements of physiological events in living cells (e.g., cell cycle studies).  
  • Material Sciences: used for investigating polymers, coatings, emulsions, etc.

 

We operate with the Zeiss LSM 710, a confocal laser scanning microscope able to generate high-resolution 3D images of specimens with high sensitivity and low photodamage.


 

System components:

LASERs

Laser Unit

Wavelength

Maximum Power

Diode 405-30

405 nm

30 mW

Argon

458, 488 and 514 nm

25 mW

DPSS 561-10

561 nm

15 mW

HeNe633

633 nm

5 mW

 

Objectives

Magnification

Model

Immersion

10x

EC Plan-Neofluar            

air

40x

EC Plan-Neofluar

oil

63x

Plan-Apochromat

oil

 

Filter sets (ocular)

Reflectors

Position/name

Excitation

Dichroic

Emission

Green

1-38 GFP

450-490 nm

495 nm

500-550 nm

Red

2-43 DsRed

533-558 nm

570 nm

570-640 nm

Blue

3-49 DAPI

G 365 nm

395 nm

420-470 nm

 

Software

Image acquisition and analysis is performed using the Zen 2010 software that includes the following modules:

  • Z stack
  • Time Series
  • Bleaching
  • Tile Scan
  • Positions
  • Regions
Publications with our Zeiss LSM 710:

Araújo, D. et al. Chitin-Glucan Complex Hydrogels: Physical-Chemical Characterization, Stability, In Vitro Drug Permeation, and Biological Assessment in Primary Cells. Polymers (Basel). 15, (2023).
https://doi.org/10.3390/polym15040791