Tissue Curation

Handling Tissues & Dry Ice Access

Frozen tissues should be kept on dry ice that is available on a recharge basis. The dry ice cooler is located between the VLSB and LSA doors on the first floor, behind a door that goes to the loading dock and dumpsters. When taking dry ice, record the number of scoops on the clipboard sheet with your name and “MVZ Curatorial” as the lab. Tissues should be cataloged on the first floor in the liquid nitrogen facility. There is also a gray dry ice cooler in that room for storing dry ice that is left over.

Taking & Labeling Tissues

Protocols for the collection of tissues vary depending on the situation and preservation method. Review the “Guidelines for Collection of Bird Tissues” for protocols specific to bird tissues. This document includes guidelines for preserving bird tissues in liquid nitrogen, RNALater, or buffer/ethanol, as well as preservation of blood on microscope slides.

Follow the procedures below for labeling tissue vials. It is important to leave some space at the lower edge of the white label on the vial, because that will be covered up with the barcode (if there is writing, it will be hidden by the barcode).

Labeling Tissue Vials

Creating Barcoded Box Positions

Barcodes in Arctos are organized in a hierarchical system. Each tissue vial gets a barcode that goes into a position in a barcoded box. Once the tissues are ready to install into a LN2 freezer, the barcoded box gets scanned into a barcoded slot in a barcoded rack in a particular freezer. More details on installing tissues is provided below.

Cataloged and uncataloged MVZ tissues go into barcoded polycarbonate boxes with a 9 x 9 grid. Official box barcodes have the format “MVZ1234” where the four-digit barcode number follows the institution acronym. Box barcodes are already entered into Arctos, but positions must be created for the box before tissues can be scanned into position in the box. The same procedure for MVZ barcoded boxes applies to temporary barcodes that might be created for uncataloged tissues (see below). Follow these steps to create barcoded box positions:

  1.  Search for the box barcode in Arctos by entering or scanning the barcode into the “Barcode” field in Find Container “(Manage Data —> Object Tracking —> Find Container ).
  2. Click on the little box to the left of the barcode to get details for that container.
  3. Click on “Edit this Container” on the right side of the screen.
  4. Under Dimensions, enter width=13, height=5.3, length=13.
  5. Under Positions, select “Magic” —> 9 x 9 freezer box.
  6. Hit “Save Container Edits.”
  7. Exit out of that window, and search on the barcode again in Find Container.
  8. Click on “Positions” on the right side of the screen.
  9. Click “Create all new positions” at the bottom of the screen.
  10. You will now have a box with empty positions that you can use to start scanning barcoded tissue vials.
Finding box barcodes in Arctos.
Editing barcoded box to specify box configuration in Arctos.
Creating new box positions in Arctos.
Newly created positions in barcoded box.

Tracking Uncataloged Tissues

Each collection tracks uncataloged tissues in different ways. Tissue vials are treated as containers in Arctos, where they are identified by both a barcode and a label. The label is intended to correspond with the original information (e.g., preparator’s number) written on the vial. When the MVZ tissue collection was barcoded in 2009-2013, the barcode was entered into both of these fields by default. Tissues may be tracked by either the barcode or the label.

The following guidelines describe the procedures for tracking uncataloged tissues. 

Cataloging Tissues

Assume that tissues exist for an accession unless told otherwise by the collector or donor. Data on tissues should be obtained from the field/prep catalog. However, because collectors/preparators may be inconsistent in annotating tissue information, tissues should be inventoried before adding them in Arctos.

All tissues should be cataloged at the same time as voucher specimens. If the accession includes a mixture of MVZ-vouchered and non-vouchered tissues (including vouchers deposited in another institution), the non-vouchered tissues should be integrated with the vouchered material when cataloging.

Tissues are treated like any other specimen part when cataloging. Tissues may be added either in the Arctos data entry screen (before the data have been loaded), in a bulkloader spreadsheet, or after the data have been loaded.

Instructions for Arctos

Labeling Tissue Cap Inserts

Nalgene caps for standard MVZ tissue vials accommodate color-coded inserts that are used to label the collection. The collection is color-coded as follows:

      • Birds – white
      • Mammals – yellow
      • Herps – green

After specimens and tissues have been cataloged for an accession, tissue vial cap inserts should be labeled with the MVZ catalog number and the collector initial plus number using a “Cancer” pen as follows:

      1. Write the MVZ number (e.g., 191967) just below the semi-circle hole in the insert
      2. Draw a line below that number
      3. Write the collector’s initials and number below the line (e.g., DRW 174)

Because the collector’s initials and number are written on the vial, adding this information on the cap insert allows matching of the cap and correct MVZ number with the vial in case of inadvertent mixing.

If there are multiple vials for a given specimen, add a single dot above the hole for the first sample, 2 dots for the second sample, 3 dots for the third sample, and so on. If the multiple vials have different preservation methods, the first dot should be given to tissue that was preserved frozen (e.g., in liquid nitrogen, if relevant) and subsequent dot(s) to other samples with a different preservation method (e.g., RNAlater). Do not add any dots if there is only one vial per specimen.

If there are multiple inserts to write, it’s best to do them all at once before inserting then into the vial cap. To keep them in order, use wide packing tape and put a large piece that’s rolled to stick front and back onto a piece of cardboard (e.g., and old broken skin tray or any other cardboard). Stick the inserts onto the tape in the order of the cataloged samples. 

When ready to insert them into the vial caps, find the correct vial (make sure it matches the data!) and align the insert with the writing on the vial. The orientation of the writing on the cap insert should be parallel (more or less) to the writing on the side of the vial, so that it is easy to read without having to rotate the vial. Carefully put the insert into the indented part of the cap, making sure that it goes down evenly so that the insert is flush with the vial cap all around. Use forceps (or any other similar tool) to remove ice from the inside of the cap before adding the insert, if necessary.

Installing Tissues

Tissues should be installed into the collection soon after they are labeled with the MVZ number. All tissues are installed in one of the three MVZ liquid nitrogen tanks on the first floor of VLSB (Freezer 1 – Herps; Freezer 2 – Birds; Freezer 3 – Mammals). Most tissues are housed in 9 x 9 (81 position) polycarbonate boxes that are labeled with a barcode that consists of “MVZ” and a four digit number (e.g., MVZ5979). Boxes without grids also may be used for large vials or odd tissue containers (e.g., carnivore tissue in tin foil).

Follow these steps when installing tissues into the collection (see figures):

    1. In Arctos, go to Manage Data –> Object Tracking –> Find Container.
    2. On the left side of the screen in the Barcode field, scan the barcode of the box where tissues are being installed.
    3. On the center of the screen, click on the little square box to the left of the barcode number of the box.
    4. On the right side of the screen, click on “Positions”.
    5. Place your cursor in the cell for the position where you want to install the tissue, and scan the vial barcode into that position. If the vial has been scanned previously in a different box, scanning into the new box will automatically move the tissue vial in Arctos and create an empty cell where it was previously. Hit tab to move to the next cell.
    6. Continue doing this in subsequent cells (positions) for each tissue to be installed.
    7. When done with the box, find the rack in the liquid nitrogen freezer where you want to put the box. If this is an existing box that was previously installed (and to which you’re adding more tissues), put the box back and you’re done.
    8. If this is a new box that has never been installed before, you must scan the box into the right position in the rack in order to record its location. This is done in the following steps.
    9. In Arctos, go to Manage Data –> Object Tracking –> Move Container.
    10. At the bottom in the field for “Parent Barcode,” enter the barcode on the side of the slot or shelf where the tissue box will be installed.
    11. At the bottom in the field for “Child Barcode,” enter the barcode of the box with the tissues to be installed.
    12. Hit “Move Container” and you’re done. Put the rack back in the freezer.