Loading the sample

Important If time is a critical factor for scanning the sample, you should set up the scan parameters using the procedures in chapter 12 before you place the sample on the glass platen.

Before you load the sample, determine how to position the sample on the glass platen of the instrument.

11.3.1 Determining the sample orientation

The Typhoon instrument collects data from underneath the sample. Use the following guidelines when determining the sample orientation:

• Membranes—Place the sample face down on the glass platen.

• Microplate—Place the microplate on the glass platen so that the edges of

the plate are aligned with the grid markings on the glass platen.

Note: Make a note of the grid square coordinates covered by the sample. You will use this information in the Scanner Control window (section 12.2.4).

11.3.2 Determining the sample placement

Use the following guidelines and figure 11-2 to determine where on the glass platen to place the sample:

• To minimize the scan time—

- Place the sample near the A1 corner of the grid. Each lettered or

numbered grid segment is marked by two dots.

- Position a rectangular sample so that the shortest edge is along the

numbered side of the glass platen as shown in figure 11-2. Because each pass of the scan head moves the entire length of the glass platen (from front to back), placing the longest edge of the sample along the scan head path minimizes the scan time.

• To minimize the size of the image file—

- Scan only the number of grid squares covered by the sample. If you do

not want to scan the entire sample, scan the squares that contain the part of the sample you are interested in analyzing.

- (Typhoon 8610, 9210, and 9410 only) Use the 10-µm pixel size to scan

smaller samples only. Because the 10-µm pixel size creates a large file, you can scan approximately two-thirds of the total scan area before you exceed the 1-GB file size limit for a single-channel image. For a linked multichannel image, each channel cannot exceed 400 MB. Scanner Control warns you if you try to select a scan area that exceeds the two-thirds limit.

• To record multiple samples during the same scan, place the samples close

together. Make sure the samples are the same thickness. If the samples are not the same thickness, the selected focal plane might not be optimal for each sample, which can result in a bad scan, poor image, and poor quantitation results.

• Align the lanes and rows of the sample with the edges of the grid on the

Figure 11-2. Sample placement on the glass platen of the Typhoon instrument.

11.3.3 Placing a sample on the glass platen

You can place the sample directly on the glass platen. If you are scanning a membrane, place the membrane inside transparency material, sheet protectors, or a plastic bag, and then place this on the glass platen.

You place the sample on the glass platen so that the sample creates a smooth surface on the glass. Avoid trapping air bubbles between the sample and the glass platen. Air bubbles can appear on the scanned image.

Before placing a wet sample on the glass platen of the instrument, squirt a little buffer or distilled water on the glass platen. The buffer or distilled water lowers the chance of trapping air bubbles between the sample and the glass platen.

Important Make sure you wipe up the excess buffer or distilled water. Using too much liquid can cause the sample to move on the glass platen during the scan, which could affect the quality of the collected data.

A 1 B C D E F G H I J K L M N O P Q 2 3 4 5 6 7 8 9101112131415161718192021 1 2 3 4 5 6 7 8 9101112131415161718192021 A B C D E F G H I J K L M N O P Q

Back of the Typhoon instrument

Front of the Typhoon instrument Coordinates of this grid square, A1 Top view Coordinates of this grid square, L10

To place a sample on the glass platen of the Typhoon instrument—

1. If the sample lid is not already open, push up on the lid release under the center front of the lid until the lid opens (figure 11-1).

2. (Optional) For a wet sample, squirt a little buffer or distilled water on the glass platen.

3. Hold the sample by the edges or use forceps to handle the sample. Gently lower the sample onto the glass platen starting at one edge. Do not trap air bubbles under the sample or scratch the glass.

Important Do not touch the glass platen or the part of the sample that will be read by the Typhoon instrument. Oil from fingerprints and powder from gloves, even thoroughly washed gloves, can leave a mark that affects the quality of the image.

4. Make a note of the grid square coordinates of the area you want to scan (figure 11-2). You will use this information later in the Scanner Control window (section 12.2.3).

5. Close the sample lid and press the lid down until the latch clicks. The sample is ready to scan.

chemiluminescent