The Microscopy Imaging Core operates as a Biosafety Level 2 laboratory and the microscope rooms are equipped with class 3b and 4 lasers, so all users must abide by the following safety measures:

• No food or drink is allowed in the microscope rooms at any time.
• When a laser is in operation, access to the laboratory is limited to only those person(s) necessary to perform the experiments.
• The presence in Core labs of individuals other than those performing experiments (e.g., students for an educational purpose) must be approved in advance by Core Staff.
• No alterations to hardware or software may be made by Core users. Any alterations must be approved and carried out by Core Staff. Any approved modifications to microscope configurations must be returned to the previous configuration when the work is complete.
• If any microscope component is broken or malfunctioning, users should notify Core Staff immediately. Prompt notification will lead to prompt repair.
• Damage to equipment during use must be reported immediately to Core Staff.  If reported, only the cost of repair will be charged to the associated lab.  Failure to report damage to the equipment will result in loss of access of the user to the facility as well as charges to the associated lab to cover repairs.

• All users must be trained by a Core Specialist and be approved to use the equipment.  Established users are not permitted to train new users under any circumstances.
• Refresher training may be required for any user who has not used the facility equipment for 6 months or more.
  
  Violation of these rules can result in retraction of facility privileges for the associated lab.

In order to ensure equal access for personnel of all labs, all users must adhere to the following rules for microscope reservation:

• Users may reserve microscope time through the online calendar no more than 2 weeks in advance unless first cleared with Core Staff based on experimental necessity.
• Users must include phone number (and email address) in the online reservation.
• Approval for after-hours/weekends use must be given by Facility staff and is restricted to experienced users who have demonstrated a high degree of proficiency and independence on a given instrument- at least 10 imaging sessions following training are required before a user may apply for after-hours privileges.
• Peak hours are 9am-5pm, daily.
• For use with fixed samples, advance reservations for use during peak hours are subject to a 4-hour/day maximum on all systems. Users wishing to reserve a block of time longer than 4 hours must use off-peak hours.
• The four-hour limit per system does not apply when imaging live samples.
• Users may extend their session on the day of the reservation if that time is still available.
• Users may not advance-reserve more than 12 hours of peak usage time per week (9am-5pm). No laboratory may advance-reserve for more than 24 hours of microscope time per week, not including same-day and after-hours reservations.
• Each laboratory is limited to 2 time slots a day/microscope.
• Reservation for overnight imaging may not extend into peak hours of the next day.
• If your experiment requires uninterrupted imaging exceeding the stated time limits, please contact Core Staff for approval.
• If a user is found to use an instrument during an unscheduled time or significantly beyond the scheduled time (including after hours), the user's PI will be notified, and the user's after-hours access may be revoked at the discretion of Core Staff.
• Sign-up slots may be cancelled at any time for required maintenance or repairs.
  
  Any unapproved reservations in disagreement with the rules will be deleted.

• Please start your session punctually (on regular hours as well as evenings and weekends) and fill out the log sheet.
• If a user is late to their reserved session by more than 1 hour, another user may take over the time slot after contacting the user with the reservation.
• Extreme attention must be paid to the safety of microscope objectives. Always be aware of the lens position to avoid breakage on the slide or microscope stage and always lower the nosepiece before switching objectives.
• If you finish more than 30 min early, please update your end-time on the booking and let the next user know that the instrument is available to use.
• Arc lamps must be kept on for 30 min minimum and need to cool for 30 min before being reignited.
• Check the online booking calendar at the end of your session; leave the system on if the next user is booked within 2 hr., otherwise turn it off. Keep lasers in standby mode whenever possible to prolong their life.
• Leave the microscope clean and ready for the next user. Use lens paper and cleaning solution to clean any oil/water objectives you used. Permanent damage can be caused by failing to clean objectives properly.
• Users are responsible for cleaning up the microscope tables upon completion of imaging work. Slides, coverslips, specimens, and other tools must be removed from the microscopes and the air table should be cleaned.

• To ensure efficient core operation, overbooking instrument time is strongly discouraged (see limitations in advance-booking).
• If you need to cancel your session, please delete your reserved time on the booking calendar as far ahead of the start time as possible. The latest cancellation time to avoid charges is by 5 pm on the evening preceding the reservation.
• If you cancel your session late (last-minute withdrawal), YOU are responsible for ensuring the microscope is turned off, if appropriate. The person before you will leave the microscope on if someone has reserved after them. This is particularly important if you are to be the last user of the day.
• In case of recurring last-minute cancellations/changes, the charge will be made for the entire time slot missed, including after hours and weekends, by an hourly rate.
• If you fail to cancel your reservation and don't use the system, an hourly rate charge will be made for the entire reserved time, including after hours and weekends.

Violations of Core Facility policies will be enforced as below:

• First - verbal warning from Core Staff
• Second - written warning from Core Staff, cc’d to PI
• Third - loss of user privileges until PI meets with Core Staff and Director.
• Fourth - loss of user privileges.

Please credit the microscopy facility in your publications by mentioning it in the acknowledgements as “The Microscopy Imaging Core at Louisiana State University Health Sciences Center Shreveport - Research Core Facility, RRID:SCR_024775”. This will enable us to demonstrate our importance to the research programs at the university and help to ultimately obtain additional equipment and funding critical for meeting the needs of our users. Authorship would be appreciated in case of a scientifically significant contribution, but not for routine work.

Acknowledgement may be written as:
“Confocal images were generated using the instruments and services at The Microscopy Imaging Core at Louisiana State University Health Sciences Center Shreveport - Research Core Facility, RRID:SCR_024775"

• Quantitative colocalizationopens a pdf document
• Analyze Immunohistochemistry images in ImageJopens a pdf document
• Make Binary & intensity measurements on colocalized channels (Nikon-Nis-element)opens a pdf document
• How to open a multichannel Z-stack image in Image J/FIJIopens a pdf document
• How to convert a image to TIFF format in Image J/FIJIopens a pdf document
• Create inset in an image for publication

RCF Offline 1 and 2 are free to all LSU Health Shreveport employees and outside users for image analysis purpose, and no reservation is needed. License dongles are required for full access to some of the software. Please contact Core Staff for the dongles.

For most users, the use of RCF Offline 3 and 4 cost a minimal fee to maintain the yearly service contract with Bitplane Imaris. You get free remote support from the Imaris (Bitplane) team.