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Needful Stains

Episode Three of Blinded By Science

Harrison Bae Wein 1 May 2006

www.lablit.com/article/106

Harrison Bae Wein

Editor’s note: This is the third episode in a series of nine original stories, each written in a different style, following the career of a scientist called Fluke from graduate school to Nobel Prize. These stories should be largely understandable to a general audience, but if you are a non-scientist and are curious about what some of the technical words mean, you can browse Harrison's glossary.

It came to the Kruger lab by overnight express in a sealed Styrofoam box. Jessica, a technician in the lab, met the delivery man with the usual banter, little knowing what was about to be unleashed. "Thanks," she called as he put the box down on the floor. "Strange weather we've got today, huh?"

"The clouds are moving in," he answered, moving back toward the door. "I thought it was going to be a beautiful day this morning, but it's turned into one of the darkest afternoons I've ever seen."

Jessica looked out the window and noticed the thick gray cloud cover. The weather sure had changed quickly. The delivery man left, and Jessica went over to the small Styrofoam cooler. It was dirty and dented, with several labels haphazardly overlaid, all with handwritten addresses so that it was hard to tell exactly where the box had come from. It was addressed to no one specific in the Kruger lab, either. She shrugged her shoulders and picked it up to carry to her bench. As she walked, she could swear she heard something moving inside. Puzzled, she lowered her head to the box and listened. Some kind of dry scratching noise. She couldn't understand it.

Jessica put the cooler down and grabbed a razor blade to open it. She cut the tape carefully and opened the box. The white mist greeted her with a spicy, musty scent. Jessica dug through the dry ice searching for the source of the smell, but it was a clean box, and all she found among the dry ice was a small brown 0.5 ml microfuge tube. She held it up to the fluorescent lights, but the tube was opaque, so she had to open it up. The fluid level inside was low – probably about 100 microliters of material. Marked on the outside of the tube was only "VI66" in black permanent marker, and there was no paper anywhere describing what was in the tube.

"I wonder who ordered this stuff," Jessica said aloud.

Norman, a second year graduate student, overheard her. "What's that?" he asked.

"This tube just came in the mail. It wasn't addressed to anyone, and there's no letter or anything with it. Just 'VI66' written on the tube. I have no idea what it is."

"Don't know anything about it," Norman said.

Jessica took the tube in her hand and walked down the lab hallway. She saw Carrie, a postdoc, in the biochemistry room. "Hey, Carrie, did you order a tube of something that was supposed to come today? It says 'VI66,' nothing else."

"Nope."

Suddenly Jessica felt something. It was a movement, sort of a wobbling, and she could swear that it was coming from inside the tube.

"What's wrong?" Carrie asked, seeing the puzzled look on Jessica's face.

"Did we just have an earthquake?"

Carrie shook her head. "I didn't feel anything."

"Strange." Jessica thought she must have been imagining things. She went back to the offices and asked if anyone knew anything about a tube marked "VI66."

Fluke, a postdoc in the lab, looked up from the procedure of an experiment he was carefully planning. "VI66?" he said. "I had almost forgotten about that."

"It's yours?"

"Yeah. It's an antibody I asked for a while ago. When did it come? Just now?"

"It just came in by express. No markings, no note, no nothing except this tube." She held it out to him. "Should I put it in the minus twenty?"

"No. Just put it on my shelf in the fridge. I'll take care of it."

When Jessica turned away, she felt the tube wiggle again, an almost imperceptible shift between her fingers. She paused, and Fluke asked if anything was wrong.

"No. I just thought I felt something," she answered.

"Like what?"

She shook her head. "Never mind." She walked quickly down the hallway, anxious to get rid of the mysterious tube.

After Jessica left him, Fluke sighed and longed for the days when he was still a graduate student. He was convinced that he wouldn't now be in such a rut if he still had Adoy's help. Adoy had taught him to plan carefully and think critically, but most of all to trust his instincts and do what he thought would work, even if it seemed outrageous. But Adoy was gone now, and Fluke couldn't call or e-mail him for advice anymore.

Fluke sighed again and hoped that this antibody would be the one that got him a paper.

**********

Fluke was amazed as he looked through the microscope and adjusted the focus. There they were: beautiful kinetochores showing up as glowing green circles on the chromosomes. Fluke couldn't believe his luck. From here on, it was easy. He'd use the antibody to fish out a couple of genes, sequence them, and voila: one or two or even three papers in the bag!

Fluke was excited. He went quickly through the lab, trying to temper his excitement while telling everyone to come look at what he had seen. There were only four people working in lab at the time: Jason, an advanced graduate student; Carrie; Norman and Jessica. They followed Fluke through the curtain into the microscope room. "What do you see, Fluke? What do you see?" asked Jason.

"You'll see. Trust me."

One by one they looked through the eyepiece. Carrie was first, and she gazed through the lenses in silence. "So what do you think?" Fluke finally asked her.

"Very interesting," she said hesitantly.

"Isn't it great?"

"Well, yeah . . . yeah, it is," she added, a little more animated.

Jason was next. "I don't believe it," he said to Fluke. "Where did you get this from?"

"The Barker lab."

"Wow. Very impressive."

"Hey," Norman said after the other two had left and he got his chance to look. "Nice spindle pole staining."

"There's no spindle pole staining."

"Yes there is. Look at it." Norman backed away to let Fluke look again.

Fluke took another look. "There's no pole staining," he said.

They switched again and Norman insisted. "It's there, Fluke. Maybe you don't see it because you're not looking for it, but I can tell you it's there. Can I try out a bit of this antibody myself?"

"Um . . . sure. I guess."

"Thanks. Thanks, Fluke," he said and left the room.

"So what am I supposed to be seeing?" Jessica asked when it was her turn.

"I've got some beautiful kinetochore staining. I've never seen it like this."

Jessica looked through the lens. "That is gorgeous! There's virtually no background at all. Now what did Norman say he saw?"

"I don't know. He said he saw the poles staining. I don't see anything that looks like a pole. All I see are bright kinetochores."

"That's all I see. You know," Jessica said, looking up from the scope, "I get the feeling he doesn't really know what he's talking about."

"He's only a second year," Fluke said. "Anyway, it can't do any harm if I give him a little antibody."

"You don't have that much to begin with."

"Just a little. I can spare that."

**********

In the next few weeks, Fluke had amazing success with the antibody. It stained four clear bands with very little background in his extract on a Western Blot. He was soon on his tertiary screen of an expression library and almost ready to start sequencing. But all was not completely well.

Fluke noticed that, even though he'd given Norman five microliters of antibody and used only two or three himself, the sample in his tube kept diminishing. At this rate, he would run out before his screens were done. He asked Norman about it, but Norman said he was still using the same bit of antibody that Fluke had given him. Fluke suspected that Norman might be stealing, and so he began to watch him suspiciously.

But then came Carrie's lab meeting. Carrie announced that the VI66 antibody stained microtubules. She said she had noticed it the first day Fluke had shown her his staining, and had been using the antibody ever since.

"Why didn't you tell me?" Fluke asked. "I've been wondering who's been taking it."

"It's common lab property," she said defensively. "I don't see why I have to tell you about it."

"Because I'm the one who sent away for it and my whole project depends on it."

"So does mine."

"Why don't you just get some more?"

"There is no more."

"What do you mean there is no more? Just call up the Barker lab. They'll send it to you."

"There is no Barker lab."

"What do you mean?"

"They're gone."

"Where did they go?" Fluke exclaimed, exasperated.

"No one knows. They just packed up and left. One day they're there, the next they're not. The place is deserted. They say the lab smells of a weird fungus or something. No one wants to go in there."

"Jeez."

"But there's another problem," came a voice from the other side of the table. It was Jason.

"What?" asked Carrie.

"It doesn't stain microtubules at all. It stains actin. That's why I've been using it."

"How can it stain actin?" Carrie said condescendingly. "It stains microtubules."

"In your dreams, toots. It stains actin. There must be actin in your spindles. You've been barking up the wrong tree."

"Only a brainless idiot like you would think there's actin in the mitotic spindle."

"Wait a minute," Fluke interjected before the fight could heat up further. "It stains kinetochores. You all saw it."

"We saw nothing," Jason spat. "You think everything you see is a kinetochore. You're probably looking at my face right now thinking my eyes are kinetochores."

"No, I saw it too," Jessica defended Fluke. "There was kinetochore staining."

"What do you know? You're just a technician."

"All right, all right," barked Professor Kruger, finally getting into the argument. "Obviously you all need to work out the properties of this antibody a little better. There seems to be some cross-reactivity."

"But I only get one band."

"I get four."

"I get two."

"Well, something's going on here. I think you should all get together and figure this thing out. For now, I suggest we adjourn this lab meeting. Tempers are hot and I need a cup of coffee."

At that, the meeting ended. Everyone was too angry to talk about working out their problems, and they all went back to their benches to sulk.

**********

The peace was not to last for long. Early one morning, Carrie came to lab, went to the refrigerator and found the tube of VI66 antibody missing. She searched frantically until she found an abandoned ice bucket on Jason's bench. She opened the bucket and found the VI66 tube floating alone in a pool of melted ice. Carrie opened the tube to see that there was barely anything left inside. She paused for a minute, put the tube back, then sat down and began to wait. She smiled. It was a chilling, evil grin.

Jason was the next person to arrive in lab. "Jason," Carrie called to him in a singsong voice as the door shut behind him. "I see you've been using some of my antibody."

"VI66? It's not your antibody. It's for all of us."

"But you left it out on your bench overnight. And look at the way you've been using it up. There's almost nothing left."

"Let me see." Jason came over and looked inside the tube. "I don't understand. There was plenty in this tube when I left last night."

"Oh, I understand. I understand all right. You're lying. You used it up and you lied about it."

"I'm not lying. You probably used it up and planted it in my ice bucket," he shouted. "I'm sure I put that tube away. I never leave things out on my bench."

"Well, you did this time," she shouted into his face.

"You're crazy, you know that?"

"And you're a sucky scientist," she jabbed.

Jason grabbed the first thing to hand, a syringe filled with India ink, and squirted it straight into Carrie's eyes. The black ink spread across her white pupils as she screamed and seized the first thing her left hand encountered, a used glass pipette. Ignoring her own pain, she dug the tip into Jason's thigh with all her might.

Jason looked down in horror at the blood spreading across his pant leg. Though his leg was buckling from the pain, his fury raged. As he fell onto his knees, he opened a drawer and took out a razor. While Carrie was busy blinking and trying to rub the ink out of her eye, Jason took the opportunity to swipe the razor's blade right across her stomach.

Carrie doubled over, clutching her gut with one hand but snatching another used pipette with the other hand and driving it down into Jason's shoulder. Jason howled with pain, clutching at the newly penetrated pipette, slippery with blood.

Carrie's used Jason's distraction to reach for some Parafilm® and start wrapping it around her torso as the wound continued to gush blood. After encircling her body a couple of times, she dropped the box to the floor, its trailing strip still attached to her, and looked up to see Jason taking the second pipette out. Thinking fast, she uncapped a bottle of methanol and splashed it in his face. He shrieked at the pain in his eyes and suddenly felt his hand being placed in a cold metal chamber. Carrie slammed the door of the microfuge down on his hand and held it while she started the rotor spinning. It was an old microfuge, so the safety catch no longer worked. The metal rotor started to spin quickly, the raised bumps around its slots shredding the skin of Jason's fingers and palm. One finger had been poking into a slot – it was immediately broken.

Jason started to cry and desperately felt for the razor blade on the floor with his other hand. He cut his finger on the blade, but the pain was almost incidental. He picked up the razor and drove it into Carrie's knee.

Carrie was feeling faint, and when the razor went into her knee, she started to black out. She tried to fight it, but found herself letting Jason's hand out of the microfuge and fell to the floor.

Jason withdrew his hand, took one look with his burning, blurry eyes at the shredded flesh, at his grotesque, raw finger dangling by literally a thread, and passed out as well.

**********

That was how Jessica found them, heaped beside each other on the floor in a congealing pool of crimson. She threw up right there on the floor.

When Jessica's natural response had run its course, she tried to gather herself together and called 911. She rinsed out her mouth and left the lab once the call was made, locking the door behind her. Outside, she slumped down to the floor and waited for the police to come.

Fluke arrived before the police did, and Jessica saw that he was going to enter through the offices.

"Don't go in there, Fluke," she called frantically to him.

"Why not?" he asked, coming over to her. "What are you doing out here in the hallway anyway?"

"Something horrible has happened."

"What? What's going on? Was anyone hurt?"

Jessica nodded her head and started to cry.

"Who? Who was in there?"

"Jason and Carrie," she said through her tears.

"Are they O.K.? What happened to them?"

"They're not O.K., Fluke. They're dead."

**********

The lab was shut down for two weeks as authorities investigated the incident. It was a simple lab flare-up, they concluded. People work together for too long and they get tired of each other. They get on each other's nerves and sometimes things just blow up. Professor Kruger and the members of his lab knew about the tensions that had been building, but they never imagined something like this would happen. The police captain said it happened all the time. "Just last month, a bunch of lab workers were poisoned by a postdoc at L'École de Brooklyn: professors, postdocs, grad students. These scientists," he said, "they're cooped up in these labs all day like rats. Someone's got to blow eventually."

Jessica didn't buy it. She pored through Carrie's and Jason's lab notebooks and came up with her own theory. "Fluke," she said to him one day. "I think we ought to talk." They went to the lunchroom and whispered even though they were alone. "Fluke, I've been looking through these books. They were both using the same cells and the same antibody – even the same concentration, but they have completely different observations written down in their notes."

"Interesting."

"I'm telling you, this antibody is bad. It somehow made them kill each other."

"Jessica, that's ludicrous."

"I tell you; it was the antibody. Look at their books yourself. I've spent the past week studying them. It's amazing: immunofluorescence, Western Blots – they're all different."

Fluke took Carrie's lab book and flipped through the pages marked with Jessica's yellow stickies. "All right," he said. "Let's compare this one. Carrie notes that she fixed in methanol. What about Jason?"

"Hmmm," Jessica said, looking through Jason's messy book. "There's nothing mentioned."

"Well, I know for a fact that Jason usually used a light glutaraldehyde/paraformaldehyde fix. He thought methanol was too disruptive. That would explain the difference in their staining patterns."

"Isn't that a little strange? A different fix gives completely different staining patterns?"

"It can happen."

"Then which one is real?"

"Who knows? Whichever one you want."

"Doesn't seem like science to me."

Fluke shrugged. "Let's look at the Western blots. Carrie used alkaline phosphatase."

"Jason used horseradish peroxidase and the ECL developing system."

"See? That accounts for the differences there."

"But those are only secondaries," Jessica protested. "The primaries are the same. They should give the same results."

"Not necessarily. ECL is very sensitive."

"But they didn't see any of the same bands."

"Hey – wait a minute," Fluke suddenly said, flipping the page. "This is strange. I don't see any bands on this blot at all. Maybe they've faded."

"Jason has some film here that looks like it could have been ECL, but I'm getting all my information about the bands from his notes – I don't see any bands myself."

"Well, that doesn't mean anything," Fluke waved it off. "He was so paranoid about Carrie, he may have taken the real blots home and left only the ones that hadn't been exposed long enough."

"It could be," Jessica said, "but I still think there's something strange going on here."

"Everything we've found can be explained, Jessica. There's a simple scientific explanation for all of it. I really don't think an antibody could magically cause two people to kill each other."

"Fluke, have you noticed there's been a strange musty smell in lab ever since we've gotten back?"

"Yeah. I had noticed that. I figured it must have been one of the cleaning solutions they've been using."

"No cleaning solutions smell like that. It's the same thing I smelled when I opened the box that antibody came in. I thought it was very strange, something smelling in a box of dry ice."

"That is weird."

"But there's something else, Fluke," she said ominously.

"What?"

"The wiggle." She seemed almost embarrassed to be mentioning it.

"What wiggle?"

"The tube. I could swear that, when I held it in my hand, it wiggled – as if something inside it was alive and trying to get out. It happened twice."

Fluke laughed and brushed it off: "We've all been under a lot of stress lately, Jessica, but I really think you should lay off those horror novels for a while."

**********

That night, Fluke had a dream. He found himself at the entrance to a mighty cold room. There was a voice inside that he recognized but couldn't quite place. Peering in through the little window in the door, all he saw was darkness. He opened the door and went in.

Inside, the cold room was tremendous. The light was dim – he couldn't even see the far walls. Suddenly, out of the darkness glided Norman in a lab coat and goggles. He had red colored lab tape arranged like war paint on his cheeks, and was holding a broken, bloody pipette. He screamed, "You took my antibody, Fluke! You took my thesis project! You don't deserve to live!"

Fluke looked around frantically and grabbed a big affinity column from the counter beside him. As Norman approached, he raised the column over his head and crashed it down onto Norman's skull, cracking the column. Norman collapsed, and Fluke looked down to see his facade cracked open like a freeze fracture. Underneath, Fluke saw his own bloodied face oozing with white column resin. He screamed.

Fluke woke up with a start and looked around. The centrifuge to his right was beeping; his spin was down. "Damn," he said. "I've got to stop doing these all-nighters in lab!"

**********

Fluke went home to get some sleep in his own bed. When he came back that morning, Norman was sitting at his lab bench waiting for him.

"Flu-uke," he called in a singsong voice. "I'm glad you're the first one here."

"I'm glad to see you too, Norman," Fluke said tentatively.

"I know what you did to my experiment," Norman said as Fluke approached.

"What did I do?"

"Don't play innocent with me, Fluke. I know what you did. You don't think my graduating from this place is important, do you? You've got to get your papers done. That's all that matters, isn't it Fluke? Not my stuff – I don't count."

When Fluke got close to him, Norman sprang from his chair and tackled him to the floor. He tied Fluke's hands behind his back and turned him over.

"What are you doing, Norman?" Fluke screamed. "Are you crazy?"

"Payback time," Norman growled, picking up a pipetman. He bent down and inserted the tip into Fluke's nose, then ejected it. Fluke felt a sharp sting of pain as the tip dug into his nasal membrane and lodged there. Norman loaded another tip onto the end.

"Norman, don't! Please. You're not yourself. You don't know what you're doing!"

"Oh, I know what I'm doing, all right." Norman bent down and shot the tip into Fluke's other nostril. The sharp burning made Fluke's eyes water.

Norman stood up and attached a rubber hose to the house vacuum line, then turned the vacuum all the way up. Fluke could hear the strong sucking through the tubing. On the bench beside Norman, a black ice bucket was wobbling excitedly back and forth. Fluke hadn't noticed Norman agitate it, but Fluke had other things to worry about right now.

Norman turned back toward his victim, the tubing in his hand, a devilish glitter in his eyes.

"Whad are you doing do do?" Fluke demanded, his speech impaired by the tips up his nostrils.

"I'm going to suck your brain out through your nose!" cackled Norman.

Norman started to bend down when Fluke heard a key turning in the lock. "Hep!" Fluke shouted. "Hep!"

Jessica's head popped through the door. "Norman, what in the world are you doing?" she asked, quickly assessing the situation.

Norman grabbed a squirt bottle from the bench next to him. "Don't come any closer. I've got concentrated sulfuric acid in here. Not only will it burn your brains out, but no one will want to come near you because you'll smell bad." The ice bucket on the bench was rattling from side to side with excitement.

"Norman, put the bottle down," Jessica commanded.

"Make me."

"Norman, it's over. Put the bottle down. Everything's going to be O.K."

"I have no project," Norman suddenly cried. "Fluke ruined my project. How is everything going to be O.K.?" He burst into tears and collapsed onto the lab bench, dropping the squirt bottle onto the floor. Fluke kicked it away as soon as it hit the ground, and Jessica came over to untie him.

Norman was promptly locked up. The police captain had only this to say: "People cooped up in these labs all day like rats, someone's gotta crack. This guy did a fine job of it, I'll tell you. Sure does smell in here. Can't you open up a window or something?"

**********

Jessica searched for all the VI66 antibody solutions in the lab and spent the next two weeks analyzing their activity.

At a special lab meeting she called, Jessica presented her data. She showed slide after slide of cells with no staining, and blank blot after blank blot using every developing method under the sun. Fluke and Norman (who had already been released from the hospital with a clean bill of mental health) were amazed by the data.

"What were we seeing?" Norman asked.

"The antibody must have gone dead," Fluke reasoned.

"I don't think so," Jessica contradicted him. "I think every one of you saw just what you wanted to see."

"But you looked too," Fluke argued. "You saw that staining I pointed out. Those kinetochores were bright as day."

"But you told me they'd be, Fluke. Did you ever hear of the power of suggestion? People see what they want to see with VI66. You told me what you wanted to see, and that's what I saw. Looking at it objectively this time, I saw nothing. That's what was there all along: nothing."

"I think the antibody must have gone bad," Professor Kruger cut in. "Your explanation doesn't make any sense. It's not scientifically valid. I think you two should look into it a little more," he said to Fluke and Norman.

"Uh-uh," Fluke shook his head.

"No way," Norman agreed.

"I think it should be dumped down the drain," Jessica said.

"If you want to blow this opportunity and throw your careers away," Professor Kruger said, " then go ahead. After all, I'm just the P.I. here. What do I know, right? I've only been doing research for – what, thirty years? You obviously know better than I do."

And, of course, they did. No one said anything.

"Hey," Professor Kruger said after rubbing his chin and thinking for a while, "maybe I should try it out. After all, I've been meaning to get back to the bench for years. This would be a great opportunity for me."

No one would dare to contradict him in lab meeting, but afterward, Norman, Fluke and Jessica got together and dumped all the solutions they had into the sink, running water vigorously down the drain.

**********

Throughout the next week, strange reports were heard all along the East coast. Dolphins were getting into vicious fights, and a constant flow of mauled dolphin carcasses was washing up onto the shores. No one could explain the phenomenon. Of course, the people in Professor Kruger's lab might have had an explanation, but they didn't follow the news. They were too busy working on their experiments.

Teasers for subsequent episodes of Blinded by Science:

Beauty and The Yeast. Murder, love and intrigue during a yeast biology meeting at an idyllic Northern California conference center. Fluke meets the love of his life, Monique, and they dream of setting up a lab together, but there are one or two things getting in the way of their happiness.

Apoptosis Now. Fluke is applying for a faculty position at Braggadocio University, but they have a small job for him before he can get the position. Fluke must travel to the heart of Darness, an old biology building on campus, where the enigmatic Professor Hurtz has surrounded himself with a gang of fiercely loyal students who continue to carry out his experiments despite the department's scorn.

Selected Excerpts From The Notebook of Jane Baxman. Jane is a meticulous new technician in Fluke's lab who has a slightly distorted view of her own worth.

The A-Maize-ing Maize Man: A Tale From The Corny Side. Jack, a sloppy postdoc in Fluke's lab, spills some chemicals on himself that cause corn kernels to sprout all over his body. A nasty human tomato is willing to go to any lengths to discover the secret to Jack's transformation. It's a good thing that two FBI agents specializing in the paranormal are on the scene.

The Coli. A magazine reporter finds the story of a lifetime when he learns that a postdoc in Fluke's lab has invented a prototype teleportation device. But a horrible accident ensues when the postdoc forgets to sterilize the chamber.

Back to the Past. What's Fluke doing drunk and passed out in the back room of a pub? Finally, learn the real secret to Fluke's success.