Theriot Lab Movie Collection

As seen on CNN, the Discovery Channel and NBC Nightly News


All the movies on this page were captured by Theriot lab members or collaborators. Complimentary videotapes for educational purposes are available upon request.

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"A microscopist thinks with the eyes
and sees with the brain."

        -Prof. Daniel Mazia, 1996
 
 
 
 
 

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Greatest Hits in Host-Pathogen Relationships


A collection of educational videos featuring many pathgoens. Some of were featured in the Discovery channel documentry "Intimate Strangers."
 
 

Life history of a single infecting Listeria monocytogenes
The sequence begins with the bacterium at the lower left corner of the cell, still in the membrane-bound compartment. It breaks free from the membrane and divdes several times before the movement begins. Once movement begins, the descendants of the initial infecting bacterium spread through the whole cell very quickly. Speeded up 900X.
        --Julie Theriot & Dan Portnoy
View (1.6 MB)
Listeria monocytogenes moving in PtK2 cells
These pathogenic bacteria grow directly in the host cell cytoplasm. The phase-dense streaks behind the bacteria are the actin-rich comet tails. Actin-based motility is also used in cellular motility; this cell is using it's cytoskeleton to crawl toward the lower right-hand corner.  Speeded up 150X over real time.
        --Julie Theriot & Dan Portnoy
View (1.6 MB)
More Listeria monocytogenes moving in PtK2 cells
Speeded up 150X over real time.
        --Julie Theriot & Dan Portnoy
View (2.2 MB)
Shigella flexneri associated with actin tails in PtK2 cells
These bacteria are unrelated to L. monocytogenes but move at similar rates and with similar behaviors. S. flexneri are substantially larger than L. monocytogenes and because of this the tails appear to be phase-lucent rather than phase-dense. Speeded up 300X.
        --Julie Theriot & Marcia Goldberg
View (1 MB)
Shigella flexneri with a deletion for icsA infecting cells
Mutant S. flexneri with a deletion in icsA. These bacteria invade cells normally and grow normally in cytoplasm, but do not associate with actin and do not move. Instead, they form microcolonies in infected cells. Speeded up 200X.
        --Julie Theriot & Marcia Goldberg
View (1 MB)
Candida albicans killing macrophages from the inside-out
Candida albicans, a fungal pathogen, being consumed by mouse bone marrow macrophages. After the macrophages engulf the yeast-like C. albicans, the fungus responds by rapidly growing a "germ tube." This projection eventually pierces the macrophage from the inside, killing the attacking macrophage, while the fungus survives. Speeded up 900X.
        --Julie Theriot & Julie Koehler
View (1.7 MB)
Salmonella typhimurium invading a fibroblast cell
Salmonella typhimurium invades Henle human epithelial cells. Contact of the bacteria on the host cell surface causes the host cell to send up huge actin-rich ruffles or "splashes" that engulf the bacteria as they fold back over. The bacteria are then trapped inside large vacuoles and replicate there. Speeded up 450X
         --Julie Theriot & Jorge Galan
View (700k)
 
Acanthamoeba actin-based motility
A freshwater amoeba crawls, extending pseudopods through the force generated by actin polymerization.
         --Enrique De La Cruz & Julie Theriot
View (1.4 MB)
Vibrio cholerae colonizing human cells
Vibrio cholerae colonize the surface of HEp-2 human carcinoma cells. As more bacteria adhere to the host cell surface and secrete cholera toxin, th host cells begin to pump out water and salt due to constitutive activation of adenylyl cyclase. In the intesine, the water is pumped into the intestinal lumen, resulting in watery diarrhea. Speeded up 300X.
        --Julie Theriot & Claudette Gardel
View (2 MB)
Listeria monocytogenes moving in Xenopus extract
Listeria monocytogenes bactera moving in a cytoplasmic extract. On the left is the phase-contrast sequence to show the bacterium moving. On the right is a simultaneously recorded fluorescence sequence, showing the distribution of fluorescently-labeled actin. Speeded up 60X.
        --David Fung
View (470k)
Listeria monocytogenes travelling in spirals in Xenopus extract
A Listeria monocytogenes bacterium moving in a cytoplasmic extract. On the left is the phase-contrast sequence to show the bacterium moving. On the right is a simultaneously recorded fluorescence sequence, showing the distribution of fluorescently-labeled actin. This spiralling behavior is unusual but striking. Speeded up 60X.
        --David Fung
View (360k)
Listeria "hopping" in Xenopus extract
A Listeria monocytogenes bacterium moves in a cytoplasmic extract. The bacterium appears to be making short, discontinuous starts and stops, 'hopping,' perhaps between the slide and the coverslip. Phase contrast reveals the bacterium to be always at the front of the tail, as expected. Speeded up 60X.
        --David Fung
View (350k)

 

Multiple ActA-coated beads moving in Xenopus extract
Microscopic beads (.5 microns in diameter) coated with the ActA protein of Listeria monocytogenes move in extract containing fluorescently-labelled actin just as bacteria do.  The phase image of the bead has been superimposed over the fluorescent image and appears as a red circle.  Speeded up 60X.
        --Lisa Cameron & Julie Theriot


 

 
 
 
 
 
 
 

Current Projects

Motility initiation of Listeria
How do Listeria start moving? This movie shows a bacterium's journey from no actin to a full-fledged comet tail in Xenopus extract. Speeded up 60X.
        --Susanne Rafelski & Julie Theriot
View (300k)
Actin flashes around Yersinia pseudotuberculosis expressing IcsA.
Yersinia expressing IcsA from a low-copy plasmid in GFP-actin MDCK cells exhibits rapid actin polymerization and depolymerization.
        --Denise Monack & Julie Theriot
View (250k)
New frontiers in particle tracking and image analysis.
New software specifically designed to automatically track large numbers of particles or bacteria undergoing actin-based motility shows Listeria movement from the bug's point of view. The first movie shows two bacteria (green) moving in extract with red comet tails. The latter two movies are revised to show only a view centered on each bacterium. Speeded up 15X.
        --Fred Soo & Julie Theriot
View 1 (17.4 MB)
View 2 (5.7 MB)
View 3 (4.1 MB)
Listeria monocytogenes rotates during actin-based motility.
By attaching very small fluorescent beads to Listeria, we can observe the bacteria rotating longitudinally as they move. Panels, from left to right: fluorescence image of beads, phase contrast image, beads superimposed as white dots on phase contrast. Speeded up 60X.
        --Jennifer Robbins & Julie Theriot
View (750k)
Keratocyte actin-based motility.
A fish keratocyte, recently freed from the confines of a goby scale, crawls rapidly using actin-based motility. Speeded up 30X
        --Rachael Ream, George Somero & Julie Theriot
View (450k)

 

 
Skidding motility of mutant Listeria
Listeria expressing an ActA with a single point mutation exhibit bizarre motility. The host cells are MDCK cells expressing GFP-actin. Left, actin fluorescence. Right, phase contrast. Speeded up 60x.
        --Susanne Rafelski, Pete Lauer, Dan Portnoy & Julie Theriot
View (1.4 MB)
 
A single ActA-coated bead moving in Xenopus extract
 
Here, a .5-micron bead coated with ActA moves in extract.  The phase image of the bead has been superimposed over the fluorescent image and appears as a red circle.  Speeded up 60X.
 
       --Lisa Cameron & Julie Theriot
View (140k)
Asymmetrically-coated large beads moving in extract
Microscopic beads (2 microns in diameter) shown here are asymmetrically coated with ActA by a low-angle shadwoing procedure developed by Matt.  As you can see, they move in extract containing fluorescently-labelled actin just as bacteria do.  Particles this large (approximately the size of bacteria) require asymmetric coating to nucleate actin tails.
        --Matthew Footer
not yet available
A symmetrically-coated bead breaks symmetry to start moving
A bead uniformly coated with ActA protein 'spontaneously' breaks its symmetry as it is propelled forward by fluctuations in the actin cloud surrounding it.  This phenomenon is mathematically predicted by a modified Brownian ratchet (van Oudenaarden and Theriot, 1999).  Speeded up 60X.
        --Alexander van Oudenaarden & Julie Theriot
View (170k)
Laser trapping of an ActA-coated bead with tail
A bead is manipulated via laser trap in this DIC image. The tail does not separate from the bead.  Real Time.
        --Lisa Cameron, Koen Visscher, Steven Block & Julie Theriot
View (5.5MB)
Listeria monocytogenes moving in GFP-actin-transfected cell
MDCK canine kidney cells, which constitutively express green fluorescent protein fused with actin, are infected with Listeria monocytogenes.  The GFP-actin makes study of Listeria in vacuole-dense, tall cells feasible.  Speeded up 60X.
        --Jennifer Robbins, Angela Barth , Eugenio de Hostos & Julie Theriot
View (450k)
Secondary vacuole dissolution following recipient cell uptake of a Listeria protrusion
MDCK cells which constitutively express green fluorescent protein fused with actin are infected with Listeria monocytogenes. Expression levels vary from cell to cell; the bacteria you see here are infecting a cell expressing very little GFP-actin. Some cells are also membrane-labelled with a red dye, enabling us to see when the membrane surrounding the vacuole vanishes. Three protrusion/vacuoles can be seen. Panels, from top left: actin fluorescence, membrane fluorescence, phase contrast, triple overlay. Speeded up 180X.
        --Jennifer Robbins, Angela Barth , Eugenio de Hostos & Julie Theriot
View (870k)


 

 
 
 
 
 
 
 

Prime Time Entertainment

...and just as entertaining.

Product placement of Listeria (Mission: Possible)
        Part of our ongoing campaign to sexify L. monocytogenes. Hey, it worked for Apple.
View (3.9 MB)
It's a bird, it's a plane, it's...a bug
L. monocytogenesactin-based motility in a nutshell.  Polymerization occurs at the back of the bacterium, depolymerization is uniform throughout the tail and is governed by host cytoskeletal dynamics, and the tail is stationary with respect to the cytoplasm.


View (800k)

Manipulated reality.
No one can be told what the cytoskeleton is.  You have to see it for yourself.


View (6.1MB)

 

Creepy.
Three video microscopists disappeared into a room near Palo Alto, California. One year later their footage was found.


View (5.1MB)

 

The Menace.
Bacterial invasion if the cell were Industrial Light and Magic (with a slightly more cramped budget).


View (12.7MB)

 

Dancing Cells
Upon observing moving keratocytes in the cell frame of reference, we immediately noticed striking periodic oscillatory motions: some symmetric, others side-to-side. Keratocytes donít just crawl; they dance. This music video accompanied an interpretive dance--inspired by some of the typical keratocyte dance moves--our lab performed at the biochemistry departmentís annual research conference in 2003.


View (17.7MB)