Imaging Facility

About the Biology Department Imaging Facility

We offer resources including state-of-the-art microscopes, and computers to visualize and process image data, and are available for consultation about the design of imaging experiments, and/or the analysis of image data and instrument training.

Location

The Biology Department Imaging Facility is located on the Danforth Campus in the lower level (room 40) of McDonnell Hall.

Contact

Dianne Duncan, Director | 314-935-6890

"I've spent more time than many will believe [making microscopic observations], but I've done them with joy, and I've taken no notice those who have said why take so much trouble and what good is it?" 
-- Antonie van Leeuwenhoek (1632-1723)

Instrumentation

The Nikon Eclipse 80i

The Nikon Eclipse 80i is a fixed-staged upright microscope for basic fluorescence imaging needs delivering high signal to noise ratios, which produces high-quality fluorescence images. The microscope is configured to image in transmitted light, Differential Interference Contrast (DIC) and epifluorescence modes.

Microscope Details

Nikon A1Si Laser Scanning Confocal Microscope

The Biology Department Nikon A1Si LSCM offers many features for acquisition and analysis of high-quality fluorescent and Differential Interference Contrast images. Confocal microscopy uses optical methods to remove the out-of-focus blur of fluorescence microscopy images. In widefield microscopy, not only is the plane of focus illuminated but also much of the specimen above and below this point, resulting in out-of-focus blur from these areas. This out-of-focus light leads to a reduction in image contrast and a decrease in resolution by obscuring important structures of interest, particularly in thick specimens.

Microscope Details

Facilities Sign-up Sheet

Provided by the University of Georgia's Faces scheduling system. A Faces account will be required to schedule time with the facility. Please contact Dianne Duncan (duncan_d@wustl.edu) to request an account.

Sign Up

User Rules and Fees

Microscope Fees

  • Nikon A1: $32.00/hour
  • Nikon 80i: $10.00/hour
  • Post-acquisition workstations: free
  • Training: $50.00/hour (2 hours minimum, all microscopes except Nikon 80i)
  • Consulting: $50.00/hour
  • 25% reduced fee for more than 30h/month.
  • Users will not be charged if the equipment is not functioning. Please inform us immediately if there is a problem with the equipment so that we can arrange for them to be fixed as quickly as possible. Please email the facility to have the corresponding charges canceled.

Training

  • Please consult the facility director at least a week before you plan to use the microscopes for the first time. Although we aim to help all users as quickly as possible, both the staff and the microscopes are often booked in advance.
  • Before being issued online booking access and using the Imaging Facility all users must be trained by the facility director or a representative from the microscope company. Users will not be permitted to train other new users.
  • Each user must also fill out a training request and provide a grant number for billing purposes.
  • An appointment will then be made for the first training session, after which the user will be encouraged to use the microscopes independently. Please note that you will be unable to book any microscope until you have been trained and your training validated for that particular microscope.
  • Every user must have their own account - it is not permitted for multiple users to share one account.
  • Initial training sessions will be scheduled for a minimum of 2 hours ($50/hour) and the user is required to bring a sample similar to one they will be using for their project to work with during the training session. This makes the training more specific to a given application and helps address specific sample issues.
  • Users can only work on instruments they have been trained on.

Scheduling the Microscopes

  • Sessions are limited to four hours in duration. If you require a session that is longer, please contact the facility director prior to reserving time.
  • If you are unable to use the microscope during the time that you have reserved, please delete your reservation in advance.
  • Users who do not arrive within 20 minutes of the beginning of the time block that they have reserved will forfeit their time on the microscope.

Canceling

  • If you need to cancel your session, please delete your reserved time on the booking calendar
  • If you cancel your session you are still responsible for ensuring the microscope is turned off.
  • If you are last user of the day and unable to keep the appointment please notify the previous user so the system will be shut down properly.

Using the Microscopes

  • Always reserve the microscope using the online booking calendar.
  • Please start your session punctually.
  • Turn each microscope on and off as described.
  • Fill in the microscope logbook to indicate use.
  • If you finish more than 30 min early, contact the next user.
  • Check the online booking calendar at the end of your session; leave the system on if someone is booked next within 2 hours, otherwise turn it off.
  • Leave the microscope and room clean and ready for the next user. Clean any oil/water objectives you used. Please dispose of your samples in your lab.
  • You are not allowed to install any software in the microscope computers or standalone workstations. Please contact the facility if you need any additional software or macros for your applications.
  • No food and drinks are allowed in the microscope rooms.

You are responsible for your data

  • Any images left on the computer hard-drives are unsafe and may be deleted without warning. Store your images at the end of your session.
  • Please do not leave behind copies of your files because the computers quickly get overloaded by the volume of image files produced.

Research

Research in the Duncan Laboratory

Our lab is interested in the genetic control of patterning: how a simple egg is programmed to generate the great tissue complexity of adult organisms. We address this question using the genetically tractable fruit fly Drosophila melanogaster, which has a long and successful history for generating insights into the development of complex organisms. Recently we have begun to address the role of metamorphosis in determining the final adult form of the fly, an area that has been surprisingly sparsely researched, despite the inherent fascination we all have for this process. (Almost every child has at one time or another watched a caterpillar form a chrysalis around itself, and waited with unchildlike patience for a beautiful butterfly to emerge.)

Although not widely appreciated, we have found that a great deal of patterning occurs during the pupal period, and that the steroid hormone response gene E93 regulates much of this patterning. The mode of action of E93 appears to be as a pupal-specific cofactor for many of the genes necessary to remodel the worm-like larva into the adult insect form. Recent work from other labs has demonstrated that there is a global alteration in the genes targeted by some regulatory proteins at the larva to pupa transition. We have demonstrated that E93 is directly responsible for one of these shifts in target gene response: E93 promotes the development of bract cells in the fly legs by integrating spatial cues from the Epidermal Growth Factor (EGF) signaling pathway with a temporal signal derived from the steroid hormone Ecdysone. Since mutations in the E93 gene affect most of the structures patterned in the pupa, it is likely that E93 will be found to have a role in many of the target gene response changes in the pupa, an area that we are actively investigating. This work is likely to have direct clinical relevance: there are known human E93 homologues, and human growth and maturation are regulated by steroid hormones. In addition, mutations in EGF signaling have been implicated in several human cancers.

We have for many years investigated the roles of several other genes in the development of the adult: we cloned and characterized spineless (ss), a gene necessary to pattern the adult olfactory organs (the antenna and maxillary palps). We showed that ss is homologous to the human dioxin receptor (Ahr), which mediates the toxic effects of dioxin in mammals. We are continuing to investigate the role of ss in olfaction and looking for the relationship between the seemingly disparate functions of Ss and Ahr. In addition, we have made considerable progress in elucidating the genetic circuitry that patterns the adult abdomen, a process which occurs almost completely during the pupal period. Both of these research areas are now tied together with our analysis of E93, since the pattern of the antenna, maxillary palps, and the abdomen are all disrupted in E93 mutations, and preliminary evidence suggests that Ss may directly interact with E93. Our future goals will be to determine the extent to which E93 patterns the pupa, whether other E93-like genes also contribute, and to gain insight into the molecular mechanisms by which E93, ss and other pupal patterning genes work.

Check out the Gallery below for images relating to this research.

Publications

  • Mou X, Duncan DM, Baehrecke EH, Duncan I. Control of target gene specificity during metamorphosis by the steroid response gene E93. Proc. Natl. Acad. Sci. USA. 2012. 109(8): 2949-2954.
  • Duncan D, Kiefel P, Duncan I. Control of the spineless antennal enhancer: direct repression of antennal target genes by Antennapedia. Dev. Biol. 2010. 347: 82-91.
  • Emmons RB, Duncan D, Duncan I. Regulation of the Drosophila distal antennal determinant spineless. Dev. Biol. 2007. 302(2): 412-426.
  • Wernet MF, Mazzoni EO, Celik A, Duncan DM, Duncan I, Desplan C. Stochastic Spineless expression creates the retinal mosaic for colour vision. Nature. 2006. 440(7081): 174-180.
  • Kankel MW, Duncan DM, Duncan I. A screen for genes that interact with the Drosophila pair-rule segmentation gene fushi tarazu. Genetics. 2004. 168(1): 161-180.
  • Kopp A, Duncan I. Anteroposterior patterning in adult abdominal segments of Drosophila. Dev. Biol. 2002. 242(1): 15-30.
  • Emmons RB, Duncan D, Estes PA, Kiefel P, Mosher JT, Sonnenfeld M, Ward MP, Duncan I, Crews ST. The Spineless-aristapedia and Tango bHLH-PAS proteins interact to control antennal and tarsal development in Drosophila. Development. 1999. 126(17): 3937-3945.
  • Duncan DM, Burgess EA, Duncan I. Control of distal antennal identity and tarsal development in Drosophila by spineless-aristapedia, a homolog of the mammalian dioxin receptor. Genes Dev. 1998. 12(9): 1290-1303.

contact

The Biology Department Imaging Facility is located in room 40 of McDonnell Hall on the Danforth Campus. If you have any questions or are interested in using our equipment, please get in touch.

Contact Dianne Duncan