Archive for the ‘project plans and materials’ Category

Engineering design of mobile cryobook unit in progress.

November 30, 2010

Design: David St.Onge

Design David St.Onge


Design: David St Onge

Design: David St. Onge

Engineering of the portable cryofreezer

November 26, 2010

Image and design: David St Onge

image and design: David st Onge

image and design: David st Onge

005-RACK
006-INTPANEL

Since there are multiple collaborators and people curious about the making of this piece I thought I’d keep posting the technical development and engineering side of the project . So here’s the latest update….
David St Onge (engineering collaborator) is busy flushing out the details for the portable cryobook unit which will be powered by dry ice when not hooked up to the -80 freezer. David and Jean-Francois made also arrangements with Multiver, a quebec company, that is generously donating the special window glaze needed for the unit. Once the samples arrive, the prototype will be build based on the final specs.
For my part, I have been on the phone with -80 freezer companies and purchasing services at Concordia trying to get the best deal for a -80 freezer unit as quickly as possible. I ended up going with the So Low brand mentioned in the previous post. Hopefully it wont get held up in custom at the Canada/US border and arrives here in the next two weeks.

Design and plans

November 24, 2010

The designs are now in progress for the cryobook archive display unit that will be featured in the upcoming show at the ScienceGallery and SymbioticA 10th Anniversary Show in January.

Here is a working design by David St.Onge that factors in the feasibility report created by Benoit and Jean-Michel  and the concept of the portable  and mobile library/archive.

The design follows the form of early portable libraries used in the 1800’s for transporting books to lighthouse outposts in the Great Lakes regions (USA and Canada).

For more background on the use of portable libraries see the following link (where these images are borrowed from) here.

The portable unit (maintained by dry ice ) will parasite off of another portable -80 freezer feeding off of electricity. This is a standard unit used for biomedical samples and specimens that will be integrated as part of the installation. (It weighs approx. 36 pounds, and the outer dimensions are 27.5” x 13.75” x 18.25” .

Building the Cryobook Archives display unit

November 10, 2010

It’s been more than a year since my last post. Hard to believe so much time has gone by. I’ve been working on developing a portable -80 cryogenic freezer unit to display the Cryobook Archives–and it has taken more time than expected. This summer I began a conversation with David St. Onge, an engineer and member of NXI Gestatio.  He agreed to take on the engineering aspect of this project and has since overseen the development of a feasibility report for the freezer display unit.  Jean-Michel Dussault and Benoit Allen, both at the University of Laval, have worked  on formulating the calculations and researching materials needed for developing various prototypes. (They also took the video posted above).

The inspiration for getting the freezer unit done soon is due to the fact that the Cryobook Archives will be featured in the exhibition curated by SymbioticA at the ScienceGallery January 28-February, 2011. We dont have much time to complete the unit and test it before I have to pack it in my suitcase and take it with me to Dublin. More to come.

limited edition cryobook series

July 4, 2009

allbookstwobookshumanskinfatbookbooksinfreezer

last series of books

July 4, 2009

The last series of books turned out in an unexpected way. The stamp imprints stained white instead of blue! Stuart said it probably had to do with the proportions of chemical mix. So now the imprint designs are white and the background is bluish/brown. In other words, I screwed up on mixing  the 4CN solution. But, I am  happy with the results. The designs on these books are by John Greyson (left) and Vincent Chevalier (middle). This is what the stamps look like.(My design is on the right).

customstamps

here are the imprints as seen on the book sculptures:

lastseries

vincentbookClose up of Vincent’s design on front cover. (Imprint on pig skin).

backbookimprint of both designs on back of book. (also imprint on pig skin).

First series of cryobooks

June 28, 2009

meandbookbooksgoodclosebook

stampimage

The first two cryobooks are finally done after two months of intensive development.

These books feature a custom designed stamp print by myself. (see above).The image imprinted within the skin of the books is a new sign for HIV.  (The red ribbon is in need of an update!). The rendering is not merely a representation, but a bringing into being  living signs.  Each stamped image is made visible by Lentivirus and HaCat cells that have been stained.

The next series will feature new designs for HIV  by Canadian performance artist Vincent Chevalier and Canadian Filmmaker and HIV/AIDS activist John Greyson.

Updated protocol

June 25, 2009

stamppages

I am now finalizing the latest version of the “Viral Tattoos recipe” originally used for the Living Viral Tattoos project (developed in collab with Dr. Stuart Hodgetts). I realize that not everyone cares about the technical application or development of this project– so if youre one of those people just skip this post. Conceptual and philosophical meanderings can be found in my upcoming phD dissertation, exhibitions and future publications.

This version of the protocol has more detail and some corrections! I am also finalizing a co-authored (hybrid) technical paper outlining the first process used for the Livign Viral tattoos project with Dr. Hodgetts, Dr. Jill Muhling and Maria Grade Godinho that will be submitted for publication at some point soon.  It’s written in such a way that it falls inbetween the cracks of a science paper and a technical/methodological art production paper.  The intention is to promote open source sharing of the wet protocols in a similar way that artists working in open source software might share programming code.

New Viral Tattoos Recipe (modified for Cryobooks Archives) June 2009
For visible expression of transfected cells on epidermis

Determine Multiplicity of infection (MOI) based on amount of viral vector to cell culture ratio. We used 1.0x 10 4 Transducing particles (TU) per ml and then divided accordingly to test for best MOI rate.

1. Suspend cells grown in flasks with 5 ml Trypsin.
2. Add DMEM medium with 10% FBS
3. Count cells
4. Add 1x 10 (power of 4) cells to 1 ml cell culture in each 24 well dish. (Optional – add Poly-L-Lysine for stronger adhesion of cells on plates).
5. Put dish in incubator overnight (12 hours)
6. Check that cells are semi-confluent (60 % is best).
7. Put fresh medium (1 ml) in the dishes. Put 2ml of medium in first dish (Nat)
8. Take 10 microliters of lentivirus and put in first dish.
9. Pipette 1ml into second dish—1ml into the third to the sixth well (to dilute the viral nate. (1/2, 1/4, 1/8, 1/16, 1/33)
10. Put 1.1 ml in the second dish. Take another 10 microliters of lentivirus. Take 100 microliters into each of the other six dishes. ( 1/10, 1/100, 1/000, etc)
11. Add polybrane ( 8 ug/ml)
12. Seal dish with parafilm and put in incubator for 4 to 12 hours.
13. Replace medium (without FBS)
14. Place in incubator for 12 hours.

Option: Skip the cell count and hope for the best. Add as much or as little quantity of virus as you feel like to the cell culture.

Prepare skin- Wash in PBS. Cut sections with sterilized scissors prior to suspending viral cells. Place skin samples into dishes and place in -20 freezer for 15 minutes (varies depending on thickness of skin and skin type). Take out and stamp.

15. Remove medium from dishes
16. Suspend virus and cells with trypsin (fill bottom of dish) for approx 2 mins or until you see cells lifting.
17. Add medium

Pipette cells onto the surface. (Separately each skin sample with one cell dish to determine MOI with microscopy, if desired).Try to keep the concentration of cells in one area for dense expression of immunostains. (Ie. Bluer colour concentration).

Immunostaining pig and human skin (Stuart Hodgetts)

1. “Seed” skin with cells and virus in about 1ml of medium – leave for 30 minutes to two hrs. Check to insure the stamp design is still visible.
2. Fix tissue with 1:1 acetone:methanol (volume:volume or v/v) for 5-10 mins 2:1 PBS Then block in 10% FBS (V/V) and 1% BSA (V/V or in 2:3 1AB diluted in Block). (The block was added into the primary antibodies).
3. Incubate with primary antibodies for 30 mins at 37°C
4. Rinse 3x with phosphate buffered saline (PBS) or medium (serum free)
5. Incubate with secondary antibodies for 30 mins at 37°C
6. Rinse 3x with L-15 (or medium without FBS)
7. Rinse 3x with PBS or medium (serum free)
8. Develop with 4-chloro-1-naphthol (4CN) chromagen solution until desired staining is achieved
9. If required, rinse and leave submerged in tap water to intensify 4CN staining

Primary antibodies:
Thy1.1 (Mouse) diluted 1/400 (10 microliters: 4 milliliters)
Fibronectin (Rabbit)
Laminin (Rat)
=together mixed with PBS to 22 milliters.

Secondary Antibodies:
Goat anti-Rabbit Ig – horse radish peroxidase conjugated at 1/2000 in PBS
Donkey anti-mouse Ig – horse radish peroxidase conjugated at 1/2000 in PBS

4CN Chromagen:
20mg 4CN dissolved in 20ml methanol = Solution A
50mM NaCl/Tris-HCl solution = Solution B
Mix 1:5 solutions A:B then add 0.1ml hydrogen peroxide (few drops)mix again
(.2m NaCl/50 mM Tris)
Use 50ml

Additional notes on skin:
Pig skin is tough and the cells need to be pipetted in one area in order to render colour. The cells expresses more easily in the fibreblasts found in the dermal tissue of both human and pig skin.
It is recommended to submerge both types of skin in tap water for at least two days for more intensity of staining

tweeking the recipe and hours of cooking

June 17, 2009

protocolhell

Here is a shot of the new recipe/protocol in process. (Above). The solutions used for the immunohistochemical staining process after the mess has been cleaned up. (Below).

antibodies

Today was a full day and night in the ‘kitchen’ cooking up the pig skin with virus.
This time I finally figured out how to mix the antibody solutions properly thanks to Stuart (my collaborator for Living Viral Tattoos and  a research scientist based in the Anatomy and Human Biology department). Besides getting the skin stamped, transplanting the viral host cells and incubating them (aka cooking them in the oven), I was measuring and mixing solutions for hours. The non-stop pipetting action and washing the skin with PBS is mindnumbing work.

The kitchen metaphor is fitting for this process. Last week the human skin  puffed out after it had been in the incubator for four hours, losing its stamp shape. Megan, a  SymbioticA regular, saw the  bloated skin pages and commented that they were overflowing souffles.  I’m trying to created more of a ‘pancake’ so that the stamp designs hold in the viral host cells as long as possible.

Here is a short video clip of ‘cooking’ the pig skin with HaCat cells, Lentivirus and primary antobodies in an incubator.

transfectedThis photo is not in focus but here you can see how the skin looks after incubation. The stamp imprint is visible apart from the rest of the skin due to a reaction between the viral Hacat cells and the pig skin’s epidermal cells.  If the immohistochemical staining process works, this area will turn blue in the shape of the stamp design.

Preparing more skin pages

June 16, 2009

stackofpigpagesAfter the book making workshop I am inspired to try to cut pages from pig skin again. This time I went to a butcher shop near the university and bought pig skin at four times the price charged by the locals in Chinatown.  Everything was clean and pretty in the butcher shop. The lighting was low and the meat highlighted. I always feel like I’m participating in a gigantic lie when I buy animal meat and skin in shops that smell like spice and herbs.

scoringThis pig skin is much more leathery than the last patch, and I think it will work well with the stamp.  I was able to score the skin to make different shapes this time. With the human skin it was impossible to try to define any shape or form whatsoever. I  used the surgery blade like an exacto blade ( or a Stanley knife as its called here). Also, I was able to use bookmaking techniques with the pig skin (like the kettle stitch) but I end up using surgery techniques (like the surgeon stitch) with the human skin.