Confocal laser scanning microscope

 

Introduction
 

    The Confocal Laser Scanning Microscope Core Facility is located in rooms 401-3 and 110 at the Institute of Cellular and Organismic Biology. In addition to acquiring Z-stack images with optical sectioning, diverse applications are available on these systems, including image stitching, live imaging, Forster resonance energy transfer (FRET), fluorescence recovery after photobleaching (FRAP), fluorescence lifetime imaging microscopy (FLIM), and spectral detection. The facility provides experimental consultation, training in imaging acquisition platforms, and image analysis software packages. Access is typically reserved for ICOB-affiliated researchers. Please contact the Core staff for details.

Services include:

  • User training for confocal laser scanning microscopy.
  • User training for advanced applications.
  • User training for image analysis.
  • Customized macro editing.
  • Experimental consultation

Equipment available:

  • Zeiss LSM880 with Airyscan, an inverted microscope with cell cultivation chamber and motorized stage.
  • Leica TCS SP5 AOBS with multiphoton laser, an upright microscope with a motorized stage and FLIM system.
  • Leica TCS SP5 AOBS, an inverted microscope with cell cultivation chamber and motorized stage.
  • Workstations for image processing
 
 
Equipment Manager:Peggy Hsu   Tel-1531  
 

Zeiss LSM880 with Airyscan

    The Aiyscan is composed of 32 detector elements, which collect the fluorescence signal with positional information. After processing the position-coded signals by linear deconvolution, resolution in all three spatial dimensions may be improved compared to traditional laser scanning microscopes (resolution: 140 nm in x- and y-directions, and 400 nm in z-direction). Please contact the Core staff to arrange a training session before using the Airyscan.

Objective list

  • Plan-Apochromat 10x/0.45
  • Plan-Apochromat 20x/0.8
  • EC Plan-Neofluar 40x/0.75
  • LD C-Apochromat 40x/1.1 W
  • Plan-Apochromat 63x/1.40 Oil

Applications

  • Fluorescence
  • Cell morphology
  • Tile scan
  • Live cell imaging
  • FRET
  • FRAP
  • Resolution-improved Airyscan

Control software

  • Zen Black

 

Leica TCS SP5 AOBS multiphoton

     Objectives with long working distance are equipped on Leica TCS SP5 AOBS multiphoton system. Please book this system if you are attempting to acquire the signal from larger embryos or organisms. With the installation of the pulsed multiphoton laser, the fluorescence lifetime measurement and the second harmonic generating signal can also be detected through this system. Please consult to Core staff for the further training course arrangement.

Objective list

  • HC PLAPO 10x/0.4
  • HC PLAPO 20x/ 0.70
  • HCX PLAPO 40x/0.85 corr
  • HCX PLAPO 63x/1.4 oil
  • HCX APO L 10x/0.3 water
  • HCX APO L 20x/0.5 water
  • HCX APO L 40x/0.8 water

Applications

  • Fluorescence
  • Cell morphology
  • Tile scan
  • FRET
  • FRAP
  • FLIM
  • Second harmonic generation

Control software

  • LAS-AF

 

Leica TCS SP5 AOBS

    Leica TCS SP5 AOBS has been used to collect data for dozens of papers in the past ten years. The system is still able to produce publication quality images. Confocal beginners are recommended to use this system first.

Objective list 

  • HC PLAPO 10x/0.4
  • HC Plan Apo 20x/ 0.70 Ph2
  • HCX Plan Apo 40x/0.75 Ph2
  • HCX PLAPO 63x/1.4 oil lamda BL

Applications

  • Fluorescence
  • Cell morphology
  • Tile scan
  • Live cell imaging
  • FRET
  • FRAP

Control software

  • LAS-AF
 

Amira workstation

 

Imaris workstation

 

MetaMorph workstation

 

LAS-AF workstation

 

Zen workstation

 
Policies Manager:Peggy Hsu     Tel-1531  
 
 
  • Access is typically reserved for ICOB-affiliated personnel.
  • Individuals must have received training and certification as independent users before using equipment unassisted or outside of regular business hours.
  • The Imaging Core Facility is open 24 hours per day, 7 days per week with access control.
  • Only certified users may apply to access the confocal or light microscope rooms.
  • The reservation allows scheduling up to 13 days in advance.
  • In order to allow us to track and bill for utilization, all users must make reservation and sign in online prior to each period of use. 
  • If you are more than 20 minutes late during high-use periods, your time slot may be assigned to another user.
  • If you overbook time (i.e., book three hours and only use one hour), please sign out online. The booking system will send a notice to other users and release the time slot for booking.
  • Please cancel your reservation if you will not use facility. Otherwise, your booking account will be suspended for one month.
  • All users must clearly sign the work log with user name, PI name and the usage time.
  • If you have any questions or encounter trouble with the equipment, please contact the Core staff immediately. Replacing any accessories and changing the settings in the control software is not allowed.
  • All users are obligated to keep the facilities clean. including the environment and all objectives (especially the immersed objectives).
  • No drinking and eating are allowed in the confocal and light microscope rooms.
  • Experimental consumables and packages of food are not allowed to be left in the confocal and light microscope room, including in the trash can.
  • All users must use their own badges to enter confocal and microscope rooms.
  • Anyone who violates the above policies will have their access to the room revoked and their PI will be informed of the violation.

 

Pricing

 
 

 

 

Extended use

Weekdays 9 AM - 6 PM

$ 200 / hr

$ 50 / 15 min

Nights and weekends

$ 100 / hr

$ 25 / 15min

 

Billing

  • All users must clearly sign the work log with their user name, PI name and the usage time for billing purposes.
  • Costs will be charged hourly and billed monthly.
  • Every 15 minutes of extended use will be charged.
  • No charge will be levied for less than 15 minutes.

 

Acknowledgement of the Core Facility
 

All users of the Imaging Core Facility should acknowledge the facility in publications and presentations that include data generated in the facility. Please also acknowledge the macro editor who writes the customized macro or script. We also appreciate receiving an electronic version of publications that include data gathered in our facility.


Acknowledgment template

We thank the Imaging Core Facility at the Institute of Cellular and Organismic Biology, Academia Sinica, for the technical support in image acquisition and analysis.

 

 
Usage protocol Manager:Peggy Hsu    Tel-1531  
 

Service object
   ICOB members

Apply for access

  1. Contact the Core staff to arrange a training course.
  2. Contact the Core staff to become certified as an independent user.
  3. Fill out the application form. (Download)
  4. Submit a hard copy of the application to the computer room.

How to use

  1. Make a reservation online. The reservation system allows scheduling 13 days in advance. (Link)
  2. Log in online before use.(Link)
  3. Fill out the work log on the desk.
  4. Log out online after use. (Link)

Hours

    24 hours per day, 7 days per week

 

 
Contact us  
 

Peggy Hsu, PhD
Phone: 886-2-27871531
Email: peggyschsu@gate.sinica.edu.tw

 

 

 

 

 

 

128 Sec.2, Academia Rd, Nankang, Taipei 115 Taiwan, R.O.C Tel: 02-27899515 Fax: 02-27858059

icob@gate.sinica.edu.tw Copyright © ICOB 2013. All rights reserved.