Danika Allaire-Degarris 7055330 Lab 6 (Chapter 8): The Retina (Graded out of 50, worth 5% of final grade) This lab consists of two parts and is done in the lab or at home. Part 2 requires the assistance of a partner. Equipment - A computer with a web browser that is Java enabled (almost any modern computer will have such software) - A ruler - A printed copy of the last page of this lab, which contains the stimuli need to do Part 2 (Provided by us) Handing In 1. Visit the course website 2. Attach your completed lab 6 Background In class we discussed the anatomy of the retina. Here, we will explore to aspects: the receptive fields of retinal ganglion cells and the blind spot. Recall that retinal ganglion cells have centresurround receptive fields. This was discovered using single-cell recording techniques. You will be doing a simulation of the same kinds of experiments here. As for the blind spot, we do not normally notice it because the brain “fills in” the space where it would normally appear. In part two of the lab, we will look at this phenomenon. In preparation for this lab, you should read Chapter 8, sections A1 and B1. Danika Allaire-Degarris 7055330 Part 1: Mapping Receptive Fields Researchers map out receptive fields by placing micro-electrodes in a neuron then searching the area of a sense organ for a zone that affects the cell’s baseline firing rate. Here, we will simulate that activity using a Java Applet. Procedure You will be acting as a researcher and technically the computer will simulate being the participant. Go to the following website: http://tinyurl.com/6tzd9ta After a moment, you should see a window open up with a blue background. There is a large dark area on the left and a set of axes on the right. At the bottom are buttons labeled “+”, “-“, “Remove Last”, and “New Cell” (and also “Show Cell” but we don’t use it). The axes on the right show the baseline-firing rate of a simulated retinal ganglion cell. It is currently firing at a low but non-zero level. The dark area to the left is a simulated patch of retina. You will see a white circle in the upper left corner of that area. This represents a lift stimulus you will use to map out the receptive field of the simulated retinal ganglion cell. Click and drag to move it around (or, use the horizontal and vertical sliders beside and below the dark area). As you move it around the simulated patch of retina, you will see the level of activity of the simulated retinal ganglion cell change. For some areas, the retina ganglion cell activity level drops below baseline (i.e., inhibitory areas), for others it jumps far above baseline (i.e., excitatory areas). Your job is to map out the inhibitory and excitatory areas. To map out the receptive field, move the light slowly around the patch of retina while watching the firing rate of the retinal ganglion cell. When it drops, you are in an inhibitory area and should click the “-“ button. When it rises, you are in excitatory areas, and should click the “+” button. Keep doing this, adding plus and minus symbols to the retina patch, until you feel you have sufficiently mapped out the receptive field. You should figure out some kind of systematic approach and follow it. You may want to review Chapter 9, section B to see what shape the receptive field is likely to have. Once you are satisfied you have mapped out the receptive field, do a screen capture. If using Windows, press “Alt - PrntScrn” to do this. This will place a screen image in your clipboard. Paste this image into this document (Control - V). Once pasted crop the image to contain only the receptive fields. Danika Allaire-Degarris 7055330 If using OS X, press “Shift - Control - Command - 4” and select the area to capture. This will place an image on your Desktop. Drag this image into this document. Go back to the Java web application, and click the “New Cell” button and repeat the procedure. Perform the above steps until you have mapped out a total of 4 retinal ganglion cells’ receptive fields, copying and pasting each map below. Paste your three retinal ganglion field maps below RC 1 RC 2 RC 3 Danika Allaire-Degarris 7055330 RC4 Danika Allaire-Degarris 7055330 Discussion 1. Compare and contrast the three RF’s you mapped. Were they the same shape? The same size? Were they arranged as you expected? The three RFs in the experiment showed a relatively uniform size and shape across the two type of ganglion (ON/OFF and OFF/ON). Based on the predictive shape and nature of the two cells I was able to easily map out the ganglions after I discovered the pattern of the first receptive field. Of course the only true differences were the locations of the inhibitory and excitatory receptors (whether on the outer ring or the inner ring) 2. What kind of systematic approach did you take in moving the light stimulus around when you were mapping out the receptive field? Why? At first I tried scanning the fields from the top right to the top left (as if reading a page from a book) but once I mapped out the receptive field and found the circular patterns I quickly changed my approach. The systematic approach I developed included moving the light stimulus from the right of the field towards the middle until I found the first receptor location. From there I would pick a location nearby and drag the light stimulus from a location slightly closer to the edge of the field towards the middle until I found the height of the excitatory/inhibitory response. I continued in a circular motion around the field and repeated the process within the outer “ring” until I mapped out the opposing receptors in the middle of the field. Once I established my process the following receptive field maps came quickly and neatly. 3. Did the cell ever reach its maximum possible firing rate? Why or why not? What characteristics of the stimulus would you need control over to elicit a maximum response? The cells in the experiment never reached their maximum potential firing rate because of the nature of the stimulus presented. A small white light of average intensity is not the proper characteristics to evoke such a strong firing rate. What must be considered is the location, wavelength, and size of the stimulus. According to the text (Chaudhuri, 2011) the location with the least amount of rods—and thus the higher threshold for light due to increase in density of photoreceptors—is the fovea. As for the wavelength, scotopic vision is the best for detecting the presence of light stimuli as it functions in environments with low light conditions. In these conditions, light with lower wavelengths (often presenting in colours such as green and blue) are seen as brighter and so generate a greater response in the eye to the presence of light. Finally, the size of the light placed on the ganglion can easily be accommodated for by making the light small enough to target only the excitatory receptors so that the opposing inhibitory receptors do not cancel out the electrical response. Danika Allaire-Degarris 7055330 Part 1: Mapping Receptive Fields In class we discussed the fact that where the optic nerve leaves the eye (i.e., the optic disc), there are no photoreceptors. This is, therefore, a “blind spot” in your visual field, where you do not see anything. In this lab you will, with the help of a partner, map out your blind spot and make some observations of what it does to your vision. Equipment For this part of the lab, you will need a method to take notes, a ruler and the last page of this lab handout, which contains the stimuli for this lab. Procedure Practice: First, practice finding your blind spot. You may find this a little tough at first, so keep trying. See “tips” below if you’re having trouble. 1. Hold the stimulus page about 40 cm from your face with the top pair of dots horizontal, and close (or cover) your left eye. 2. Look at the left dot with your right eye (your left eye should be closed or covered) and move the page slowly towards you. You should see the right dot disappear at some point. Remember to keep your eye on the left dot or this will not work! 3. Now, keep moving the sheet towards you until the dot reappears. If you’re having trouble, keep trying! Here are a couple of tips if you are really struggling: a) Make sure you are not moving your eye. It can be tricky to keep fixated on the left dot. Just focus on that and do not worry about the right dot. Just “let it disappear”. Have you partner watch you and let you know if you are unconsciously moving your open eye. b) Try tilting the page a bit so the dots are not quite horizontal. Some people’s blind spot is a little higher or lower than horizontal. c) Try switching to the left eye (i.e., close/cover your right eye instead). It may be easier to find your blind spot in one eye than the other. If you do this, you want to look at the right dot and watch for the left dot to disappear. d) If you really can’t do it, use your lab partner’s data. If you both can’t do it, you’ll have to persuade someone to help you out. Danika Allaire-Degarris 7055330 Data Gathering, Quantitative Measurement Once you’ve practiced finding your blind spot a couple of times, it is time to map it out quantitatively. 1. First, have someone measure the distance at which the dot disappears (in cm). Typically this will be around 30 cm, but it varies considerably from person to person. Measure it 5 times and take the average. Mean distance at which dot disappears: _______31.4cm_______ 2. Next, have your partner measure the distance at which the dot re-appears (in cm) as you continue to bring the sheet closer to your face. Typically this is around 20 cm, but again varies a lot from person to person. Again, take the average of 5 measurements. Mean distance at which dot re-appears: ______25cm________ 3. Now calculate the visual angle (in degrees) of those two points using the following formula (it may look daunting at first, but don’t worry, take it easy): A = arctan (4 / X) * 2 Arctan (aka “atan” or “tan-1”) is the arctangent. Real easy to do on Excel or Numbers, simply use the =ATAN( ) function. Visual angle at which dot disappears: Visual angle at which dot reappears: ____0.253_____ ____0.317_____ 4. Take the average of these two values to get the centre point of your blind spot. Centre point of blind spot: _______0.285_______ Take the difference between these two values to get the width of your blind spot. Width of your blind spot: ________0.064_______ 5. How do your calculations compare with the average human values of 17 degrees (centre) and 5 degrees (width)? If they differ, why do you think this might be? My calculations reported higher than average values for both centre (28 degrees) and width (6 degrees). The reason for the change in blind spot angles, I believe, is due to the fact that I have astigmatism in both eyes. Should I have had normally rounded eyes my calculations would have been much closer to the average as the retinal nerve would have been properly placed in relation to the rest of the optics in my eyes. Danika Allaire-Degarris 7055330 Data Gathering, Quantitative Measurement 1. Find your blind spot as in the practice instructions above. Now, take a pencil and swipe it in front of you, through the blind spot. Try this tip first, sideways, etc. What do you observe? From swiping the pencil through my blind spot I was unsurprised to find that the parts that had travelled through my blind spot had disappeared from sight. I used the tip of the pencil at first and observed it disappear—though not entirely. What ended up happening was that the space on either side of the blind spot seemed to have fused together to create a complete image; the pencil appeared to have jumped quickly across the blind spot when I passed it through. 2. Next, find your blind spot using the second set of points on the stimulus sheet (in the middle of the page). The right one has a vertical line through it. What happens to this vertical line when the dot in the middle of it is in your blind spot? With the dot centered in my blind spot I noted that the vertical line did two things. First the parts of the line closest to the dot had disappeared along with it; secondly, the parts of the line further away from the dot stuck out of the blind spot intact. I was able to see the line clearly at the top and the bottom of the diagram with ease. 3. Finally find your blind spot using the third set of points (at the bottom of the page). The right one is surrounded by a grid. What happens to this grid when the dot in the middle of it is in your blind spot? Depending on how far away the paper was from my eye (I noted that there was a fair range of distance in which I could keep the dot in my blind spot while manipulating the paper) the grid had a few things happen to it. Further away (with the dot still in my blindspot) I could only see the higher and most left parts of the grid with about a square of depth. When the paper was close I could see the lower and most right sections of the grid. In the centre I could see the basic outline of the grid and a few of the details inside, though not very much. 4. What do these observations mean in terms of what happens to visual stimuli in the blind spot? These observations suggest that the blind spot in the field of vision is incapable of perceiving visual stimuli. While we cannot see within the blind spot, the receptor cells around the blind spot generate an image within the unperceived space that is congruent with the entire visual field perceived by the rest of the eye. Thus we do not perceive the space as being empty but a space that is unfocused and vague.