Or used to disappear so I would stay every text of the last five years I've been doing this. I can't think of a see one that is not sure. So I'm either optical or like anybody tell me it's all single so I'm writing it. I'd like to know. So that's one of the reasons are invited today speaker Richard Brown from envy a scientific side to the consultants who is a real expert in my prosperity my cross in general which is undergraduate work in chemistry at Westfield State Massachusetts and masters in forensics. There is a chemistry at Northeastern University and spent a number of years as a criminalist Orange County sheriff's office before joining and V.A. scientific career he's currently executive director so this is going to give us some real practical advice on how to go. Thanks David. Yeah we can switch him over the video you were watching was actually done and this we have a little homemade hot stage which is just a glass microscope slide. It's got an Indian tin coating on it the same type of technology that ten aircraft windshields put to it heats up. It's great for watching. Stuff Melandri crystallise So it's a what we call a poor man's hot stage. Now I have to do is drop a high definition camera on top over again. So today I wanted to resent you with the world's most powerful analytical tool. It's missing a light source and it's missing an operator or a user but most of the problems at least I deal with in a daily basis. Can only be solved with a microscope not necessary a certain essence should not necessarily a polarized light microscope. We have scanning electron microscope transmission electron microscopes for a transform infrared microscopes kind focal Remonde microscopes but this is the work course right here. So as we go through the presentation Now I'll tell you a little bit more about why we use it in the types of problems we can solve. So characterization and source identification by lighting the lights from across to be. If I can make your. You know we go. So in in our world everything we see. Well is everything most Everything is Illuminated from above the sun's above are used to seeing things in that manner. Most of what we see is in reflected light and actually be pretty weird if all of a sudden we saw everything with transmitted light. We all look a little bit different. It may not be desirable but that's the way that's what we're used to seeing. But with a microscope. We can change how all that works. So we all know we can magnify things with a microscope. That's pretty basic right. But when I only think of chemical my cross could be or problem solving with a microscope as in any type of application in chemistry. We want contrast. With a mass spec we want contrasts we want to resolve different fragments. Or the G.C. We want to resolve different. Peaks as we separate things that go through the G C. So we're looking for contrast and with a light microscope or any microscope. We're looking for magnification with the main thing we want is contrast I want you to keep that in mind as we go through. To this. So you know we got reflected light we got salt and pepper. No big deal. It looked a little bit different and transmitted like salt. It's got a single refractive index. It's crystal in but it's isotropic single refractive index pepper so organic material to actually multicellular. Anything we look at. It has different types of properties depending on how we look at it. And what it is most of my words. When I look at contamination on a surface. That drives how I actually approach if I have a polymer material fragments of a polymer I'm not probably not going to put in that any S.C.M. I'm probably going to go right to the F.B.I. are the polarized light. So as we magnify things we get more information. Transmitted light a cotton fibers. Glue pointer. Things. We can actually see. How the fibers twist. And we can actually see that the cell walls and the outside. And we've got this little lumen in the middle. So the cross-section kind of looks like. And like an infinity sign or something like that. We cross the polarizers looks totally different here. We actually have a polarizer at the base of the microscope we're putting polarized light through the sample and that's oriented east west routinely We also have an analyzer which is another polarizer in the top part of the microscope. That's oriented north south when we cross them our field goes dark. Unless there is something in the optical path that only interact with that light in this case the cottons interacting. It doesn't go to extinction. And this is just the way cotton works. It's really easy to figure out. OK this this picture's more typical of a dust sample or a contamination. We've got lots of paper fibers. Everybody see the wings yet. And sometimes it's difficult to pick up you're not really looking for that you see the insect parts. Hairy Legs. You really see the wings. Which takes a little bit of practice a lot of times you're looking at so much material at one time. It's a little bit overwhelming so it takes time for your mind to sort these things out. But again I just transmitted light would partially cross polars if we look at materials that have been coatings like mica particles with a thin layer of titanium dioxide a lot of pigments that are used on automobiles and Aviv ever walk by a car kind of looks black to get to the side and it's kind of paint or purple the color of flip flops a little bit. Back in the seventy's it was very expensive to do that but basically you've got Mica particles or some other particles that have a thin film on him. So as you walk around the car the way the light hits it changes it and it changes color. So in this case this is typical of the pigment we have on our twenty dollar bills you look at it in reflected light you see one color. Transmitted light looks totally different and these are new concepts but there are clues to me when I do my analysis when I do my examination because a lot of times Mica is present make up and a lot of folks wear make up and it gets to be a problem in manufacturing. When I when I talk to groups especially attorneys. I have a question form. And when I talk to groups especially young people. Twelve and under I asked the same question. If I had an apple in an orange. And I could shrink them down really small. Would I still be able to tell the apple from the orange shoes in a microscope. If I asked the kids at almost every hand goes up and they say yes it's a big deal. The lawyers only seen anything they think is a trick question really quiet. I don't know I don't know about that one but yes I mean soon we could do that. It's not a big deal into the to the to the light microscopy is. A lot of what we do the particles that we look at are a simple to tell apart as an apple in an orange. So just imagine for a second that these are the two particles we're dealing with immediately we see color differences we see shape differences we see differences in the surface more of ology. If we were to cut them open we see a difference in cross-section the orange should have a thicker skin in a different interior then the apple the apple have a thinner skin would be different. Inside. We do the same thing with particles. Who look at the outside. We we try to get light through and to look at the inside. Sometimes we have to cut them we have to use a moment section and sometimes we have to use a non millon section all depends on the particle the whole idea is to characterize them as best we can. And the biggest difference between characterization is and identification is a technique you used and even how much you want to pay characterization is relatively inexpensive. You get a lot of red particles they could be apples. And sometimes because some are says That's great. I know where those are coming from. But if we want to know what species apple that is actually get expensive. We may have to get a botanist and do a little more work. I don't know if they can do D.N.A. on an apple but if you can't you probably will be soon. So it's a little more expenditure. To get to the identification point. That's a very point important differentiate and different difference to make because of the cost involved in the amount of information and work you have to do. Scanning electron microscope and we fairly familiar with that. We've got an energy dispersive X. ray spectrometer attached. It really can identify anything you can tell us you can tell iron from copper and you can tell the metal alloys to some extent if you can't tell the difference. Tell me the difference between three or four three or four stainless and three sixteen which can't tell the different types apart it's not that sensitive. Plus you know any have any crystal graphic information where you got it's a black and white picture in E.D.S. spectrum. I mean you could have different detectors and you could have cathode a luminescent all kinds of different things that help you but it's not an identification technique. Poor eyes light my cross be on their hand on the other hand you have all types of optical properties you can look at and actually make an identification refractive index some particles have multiple refractive in the C.S. fire for engines. You can identify particles with mineralogist geologists have been using the techniques for years. The microscope is over three hundred years old. The polarising light microscope came about in the late eighteen hundred so that's to over two hundred years two hundred years hundred fifty years. Some way that they've been around a while. Classification and they definitely can. So first thing that usually happens is there's a problem. We don't always catch the people responsible in the act. This is rare. This is a rare thing usually the children are gone the baby powder viler bottle is gone or just this white dust. If it was my house that would be a problem. OK for the little girl who's getting the business and that this is probably a problem for her too. She's not happy. But if it happened in this school or in this room and the first person in the morning came in and this well white powder was all over the place that would be a problem. We don't know what that stuff is is it benign. Is it baby powder or did something put some anthrax or something in here is necessarily meant to be toxic or to harm people we just don't know. I was on a commission. Gees. Probably five or six years ago now World Trade Center back in two thousand and one September eleventh when the towers were destroyed by terrorists and E.P.A. eventually was trying to characterize as dust and reason is what buildings are contaminated what buildings that we have to clean up. Me being practical. I say the contamination goes from this in the Manhattan to that in the Manhattan and probably over to New Jersey and in like the audience or they didn't want to clean all of that they wanted one thing was or one thing in this dust microscopically they'll tell us it's from this event. Only answer is no. But. If we look at the dust. Transmitted light microscope. It's primarily composed composed the glass. Glass fragments glass fibers and these fibers that are kind of bent. They're actually mineral they're spun like cotton candy and used for insulation. Ceiling tiles. Just about pipe wrapping just about anything and think of the uses very inexpensive and it's a very good insulator. The problem is mostly glass. So a lot of people are breathing this dust defile different sized particles that are respirable and it's causing all kinds of problems as well as concrete dust lead particles the. The actual towers were probably the single largest architectural use of lead of any building ever made all the gutters everything were led sheeting. Led in wires. Used to connect certain devices in the building phone wire all that stuff. So we found nine years pictures sixteen electron images six. Calcium carbonate straight fibers straight glass fibers which is the insulation you probably have in your house there straight. The pink stuff the paint insulation glass fragments. Gypsum from sheet rock mineral wall which gives you these exotic shapes because it's fun like cotton candy. Lots of paper fibers as well as heavy metal particle heavy metal particles lead welding spheres. I think we found nine things that if you take them together if you find all nine of these things in the dust sample. We have a reasonable confidence that there from the World Trade Center dust. There isn't a single thing because these are very common things in your house if you have somebody and you do some sheet rocking you get is white dust all over the place going to have a lot of gypsum and calcium carbonate and maybe even some glass fears from some of the stuff they used to patch that material. So it's everywhere. Subic with this really especially in the city. So we can help the whole lot and now we didn't make them very happy but they did realize that there are two hundred dollar samples idea wasn't going to work and they went back to the old standby let's just look for its best this. And if we find a specif will clean it up and say that's related there was a lot of especially as in the buildings as well. So more typical things we do pharmaceutical folks when they manufacture a product they're very sensitive to contamination is one of our classics we get these stoppers. And we've you're all going to be the detectives a little bit here in a minute. We have the stoppers that have these black particles on them and you may you may be good enough. Right now to see what that black particle really is. So when you see it up close. It's even less likely to be make anybody happy. So all of a sudden there's a group of people that buy the stoppers packages or product with the stoppers and send their finished product on. There's also people who manufacture the stopper so we're trying to find out who's ultimately responsible for these insects getting into the stoppers any ideas. Yet you think it's to stop or manufacture some some How about the guys that are actually bottling the product. OK About fifty fifty. The rest of us and we don't care. That's OK so we have to look a little closer. So we use a scanning electron microscope course we lose color because we have electrons now and we start to see can you see the wings. And see that back there. Looks like the insects are actually embedded in a rubber. So it's highly unlikely. But not impossible but highly unlikely that the flies are actually getting crushed into the rubber stopper during manufacture of the product during the actual stop running of the vials. It's more likely that as a stopper manufacturers mourning the shape of their stoppers with their liquid stopper material silicone rubber that these flies are getting in and getting trapped in the rubber. So we voted for the stop or manufacture and it turned out to be what it was you know here's a good case of classification identification this this particular facility was in a different part of the world. And so there was some idea well we've got some pretty good replicas of these insects perhaps we can tell what they are and where they're from. Again more time more money but if it was important enough it could have been the one who we did to have some pretty good impressions of the insects. But for our customer at this point in time this was good enough. Quick answer. You know a few quick pictures and they're happy. It's not them. They'll move on to deal with a stop or manufacture. Another situation we have a. N.F.L. football player. Who is testing positive for steroids and he's not supposed to have steroids in his system. So that right there is a bad thing. He claims he doesn't take of his handlers. But he does take this protein powder muscle protein powder all the time his handlers are accused of perhaps adding something to his protein powder. So litigation happens the attorneys get involved. And now the manufacturer of the protein powder is being looked at as a source of the steroids. This means factor does not use steroids in their product they don't manufacture steroids or steroids within one hundred miles of the facility. So how do you prove that something's not there. In the manufacturing world they have something called retains So I mean your factual lot of a lot meaning a group of product on Monday and I retain a small sample of that lot. There's actually about thirty six ingredients in this stuff so analyzing it for on one hand a nightmare. You know trying to go back through and reengineer it. It's not going to happen. I just spray dried private actual It's pretty cool under the microscope. So they have always retained. So how many do we have to look at how many is enough. We looked at one hundred retains and we killed. We combed through with a stereo binocular microscope. If this is a typical field of view we estimated the number of particles in that field and we came up with something like one hundred million particles that we looked at and actually combed through. So we're trying to get some kind of detection limits. You know how do you prove a negative. Have we looked at all the product that's the only way. You know we have in science we make estimates we look at samples we estimate a population based on what we see so I mean my estimate the population is is a highly unlikely that there's any steroids in this particular product but that wasn't good enough for the lawyers. Very difficult to prove something was not there. But combing through this sample we did find this stuff which looks very different. This is the product the protein powder and this is the material we are finding and some of the Had these little big Well little white particles attached to it which weren't really little this turned out to be some kind of chinese herbal medicine and this is the steroids. So initially what happened. They sample they got hold of this. Athlete's muscle power centered off to some lab on the West Coast that's very good at detecting steroids it was at the detection limit it is huge amount of powder and it's very small amount of steroids. They couldn't detect it or if they did it was right at that attention one that not certain not certain. So I was able to go through microscopically and pick this stuff out. Piece by piece put it into a cap a plastic cap of. Put for oil over it and I think I collected when I say about fifteen milligrams of material. Send it to the lab and it tested positive for the steroids era concerned with. But there's no way to do that. I mean there's no other way to do it. You have to manually separate I thought well maybe it fluoresce maybe I just signed a U.V. light on. Take it all. Or maybe I could filter or maybe I could see if it didn't work. It was just back to this good old fashioned hard work and there's a higher main advocation look at this stuff. So you can actually take a piece of this and that I think that scales to millimeter So these are pretty big. Take a piece of that rolled out get an F.D.R. spectrum on it and it was perfect for the steroids interest. I mean steroids as a group. I don't think you can differentiate by F.D.R. but I certainly could show that there was a steroids present in it was confirmed by the other laboratory in a West Coast. He's a traditional method G.C. mass spec to identify it. That was fun. There's a couple things we can do would polarize lie. This is a court particle. This is in transmitted light would one polarizer so we still have a bright field of view refractive index at this. Rotation of the stage is very close to the liquid it's in and we use this microscope. Most of the cover most of the objectives we use that are the actual business part of the microscope that expensive part of the microscope. We have a refractive index liquid that we put the particles in and we use a cover slip. And that refractive index like that we pay for it's expensive in we know the refractive index of it at a certain temperature so we know that the refractive index shown by this particle in this orientation. Because of the lack of contrast is very very close to that of the liquid. So in the time it takes me to put the particle on the side look at it. I have one of the refractive index figured out. And then when I cross the polarizers and I see this by refrigerant I may loom. My mind goes. Maybe this is cords. You know because that index is close to one point five four and I get these in a spiral for engine colors and the fracture kind of looks like glass which with cords does it can quit. Fractured type mineral So mom a way to figure now what this stuff is included that this is kind of in the future. Is probably the last thing I'll talk about. If we've got time I'll show you a couple videos. This is a situation we had these lamps in a studio that films for television. And there's lots of these lamps in our people that get up in the rigging in movies lamps around all the time these lamps burn very very hot. So the guys who go up and the guys and gals they go up in a rigging rigging were getting very concerned about this dust that are some history to this particular location they had been in a fire and a re rebuilt it. And so there's been a lot of construction and so one point somebody said you know what I'm not messing with this stuff anymore because I don't know what that is. So we're going to stop work and so they did. So now we got to find out what it is and just by the way the first thing you notice you know this is the bald part. This is the lens par or the projector part. It looks like something's getting hot in this stuff coming from the lamp except after looking at a bunch of the rigging he couldn't be sure it was all over the place and they moved the lights around a lot but that's what that's what my first look is telling me. And sometimes at first look can be good and other times you can lead you totally in a wrong direction. So if we take some of that dust. That's on the lamp look at it under the polarized light microscope. I see two things immediately what I thought was dust really isn't. What it is we get these strings eggroll these are all tiny crystals This is poly Crystal in. And it's forming these threads or ribbons. Which tells me that this material has. Been deposited on that portion of the lamp in several events. It's it's been heated up or dissolved heated in this case as a vapor. It's the posited in its form these beautiful crystals they just keep building on themselves. So I've got some information at this point. I still don't know what it is. Actually what it is was pretty easy. So I put it in the S.C.M.. I get a picture of it. I see the same kind of thing. And I get an E.T.S. it tells me it's carbon or knocks it in. Useful because now I know it's organic as opposed to some type of mineral or some type of metal. Inside the lamp. We had more crystals a really neat This is an edge on view he got these nice little crystals all over the place. But it still doesn't tell me what it is and I still own just been looking at the lamp I took it all apart I still couldn't tell worlds coming from. So in an F.D.R. spectrum of the crystals. In. Actually this is a show it bother you got the wrong one. But that's what it is it's a acid. So it's some type of polyester material. Unfortunately just about everything they use in that studio including the daylight filters they use on the lenses were some type of polyester material were it was P T or whether it was some other polyester type material so they're all very close. So it's a OK but it's possible the lamps are heating some of these things up and they're volatile izing and getting deposited at the cooler parts in the in the ceiling. We don't know it's a little bit frustrating at that point especially after going through a lamp in the field figuring out eventually. The people who are working on this with me sent me a brand new lamp in the lamp I'd been working on. And you notice anything different. They're both black. Actually they both had marks on their manufacture in two different locations but that's OK textures a little bit different but one writes kind of shiny new and. This one been in service a while it's dull so little light goes on. It's it's got to be the pain sometimes happen into that pain sure enough I had sampled this and run a spectrum on it and I got a polyester. Which didn't help me although everything I looked at was some type of polyester just turned out. But this was still very uphill very plastic. This was very brittle. So which I would expect after I got hot and. Had been in use for a while. So I had to do a little modification here. I did take my most powerful analytical tool I had. And build myself my hot stage. And try to figure out where these crystals were coming from. So I've got a very at. We've got a tin indium slide coated slide. He just put the out put the D.C.L. put on it crossed it to the top of it's got the coating so it's conductive heated up resistivity heat it. Having it start to see now we actually left this at three hundred degrees centigrade for twenty four hours. There's three little particles of paint here. This is a glass tube or cylinder with a cover slip on top and the whole way you can see that we see the crystal starting to form after a day and believe me we tried this many many times at different temperatures for increasing the time in temperature each time. So when we finally got crystals a little party. I think where and ran down the hall he says I got the crystals and we actually have some condensation of crystals here rechristen is Asian here and we've got a boatload of them up here. And there is another microscope that it would in perfect for this application. Room on my cross could be because it will shoot through the glass and the glass doesn't bother it. So we could actually do it with the crystals on the underside and get some Ramond spectra of it. We didn't do that we flipped it over looked at them under the peel did an I.R. and then compared to what we had. And they were. Identical to the deposit on the lamp what I produced from the paint in my tree fell like acid. So my conclusion was that as these things heated up. It's the paint was breaking down into recurse the lising on this rigging which when you look at the amount the size of these lamps and the amount of dust that was produced. That's a lot of material. It was really a problem there with that pain. And that's all I have for this part. We have ten more minutes maybe. So you can do this and stop this one actually Or are we with this one. This is the hot stage set up because a lot of times it's fun. And useful to watch things rehearse the lies. I think this is T.N.T.. The explosive. So as a heated up. I can watch to Crystal's Mel here a million arrests and things like polymorphs. And if I put another material in I can look at you. Tactics this and if you have the same material you put the two tiny part of the same material melt at what should happen. They should mix. They should form a free interface. That's a good test to begin this is this is time intensive stuff you with a microscope your study and you don't you don't get your results immediately you have to study your material and then you can watch your rechristened lies and again you're looking for polymorph which are different. Crystal shapes. Actually it's a good story about T.N.T. The doctor McCrone worked on McCrone research. Since a two years ago where he found that there was an unstable mob polymorphic T.N.T. that they were loading into the shells they shot off the ships big giant shells T.N.T. is a propellant there drop and shore falling on our guys and so the bad guys. He found it was an unstable polymorph is being produced. And he's talked about that with the American Chemical Society many times before he passed away. And he wouldn't find his real quick and this. We talked a little bit about refractive index. You need. To look at the two particles left and right. I'm focusing up he see the white line the halo The Becky line the one on the left moves into the particle the one in the right moves out here to glass particles. And that much time we know what. Those two particles of glass have different for active in the Cease there from two different sources. If you will white powder and in the question is a quarter glass quartz crystal and we saw a nice picture showing the Byron is a nice colors in seconds. You can tell if it's crystal in an crystalline very powerful technique. Unfortunately takes about two years to get good at and probably five years to master so it's not something you do overnight. If you take a week or two long course you learn enough to be dangerous but you do get inspired you to like it or you hate I guess it's like computers. See if I have to put a crow is in here somewhere. And get biographer engines which scene that we can watch it in action. We're going to look at our friend courts again. One polarizer we cross them. It's dark. I apologize for that you see the by refer engines right now we know it's crystal and. We rotate the stage. It goes to extinction. No more color what that means is that's one of the refractive in the seas that particle. So we don't cross the polars So I think we'll do then a minute. See another one the upper corner another particle and now if I change my focus. I see that the bank the line moves into the particle. So I know what that rotation at that extinction point of that particle the refractive index of the particle is greater than the liquid it's in so if I know what the liquid is. I've determined not exactly what the refractive index is but I know about what it is we know we know it's greater than one point five four. So we can start a classification scheme based on that. And I won't I won't kid you it's a lot of work. But once you use to it. It's very very fast and if used the right mouse. And there's one more and I'll quit and I'll answer any questions you may have. When I'm looking for is plea a Crow ism. Pat please crows I'm the wrong color. I don't remember this one Anybody familiar with that term. Please Crowism a lot of dyes or please. Polarized light again one polar when you rotate it changes color. This is like a geologist dream because all a lot of minerals do this and they can tell. Rapidly. If a mineral is a. Type of mineral depending on how it reacts to one cross polarized beam of light by how it changes color a lot of dyes do this you see cotton fibers are dyed blue or purple and the dye will actually be plea across it may shift from blue to yellow as you turn it's not the cotton it's actually the dye that's another optical property we look at. Hopefully that made some sense or that I think I'm going to quit and see if there's any questions or anything about what you seen or anything about what I do. We have my cross to piss in here. Then we actually use microscopes electron or light. OK so I got for light my cross will get dark feel Brightfield fluorescents phase. I see. Conflict will. Yes. Can't forget that. It's really up in your mind was I don't want to implement the structure of the woods are different in paper cotton rather cotton looks very different paper can be mechanically process where they just rip. The wood apart and they'll be little pieces still stuck together of the actual cells that make up the wood or they'll be chemically process where the fibers are. Basically all it means dissolve it freeze the fibers. So the actual structure of the cellular. The cells that make up a paper fiber a wood fire very different in kind. In or are things you can see like hardwoods are different in softwoods there's a whole nother group of science paper Institute used to be around here somewhere or they were from Wisconsin. Walter rant and then a good friend of mine. He's one of the leading experts on wood. Species in paper fibers or he looks any tells what type of wood they're from so they have very unique microscopic. Structures that help identify that where the wood came from or the tree it came from or if it's recycled or how much of it's recycled. You know somebody to run it. No that's a good question. I would say I price a the I R F T A R. Because in I've got my small particles and if I if I don't really have enough. Plastics For instance if I had a Palm marriage material or a paint can I can maybe look at some of the fillers in the paint with this either mineral fillers. But as far as a resin I'd want to go to F.D.R. to help determine what the resonance for the type of polymer and again you can only go sorry. So far with that you could tell now on six from Nylon six six from six nine may be. But you're pushing it. You have to go to other techniques. Yep we have Ming do it can mean looks at a person. Now we have hoods would help of filters the negative flow. So we have a bench top with a stereo microscope in it so we open material in there. For our first look at it. So theoretically the particles won't come out into our Airstream they'll get captured by the hepa filter. And a lot of times we know. And if we don't know. Then we have to make a decision whether we want it in the lab is we're really not set up for biologicals or anthrax type things or anything that will require a high level of containment. We're careful. OK thanks very much. Thank you. Thanks for having.