Rising Pressure, page 17
“I think I have a few that worked. Juan was just explaining to me how to gauge size with the DNA size marker that I used.” Becky pointed at the gel and held up a card that had short, dark lines with a number to the left indicating how many kilobases corresponded to those positions in the marker. It was kind of like a ruler for DNA. “We were just about to take it over to the UV box to get a photo.”
“Let me know what you find. I know you’ve been anxious to start growing some protein for your binding assay.”
“I’ll keep you in the loop,” Becky said.
Their lab shared a UV box with the department, so Becky followed Juan out of the room to get a photo of her work. Addison noticed that Eleanor was off the phone, so she walked over to say hi. She hadn’t seen much of her all week because of her lectures.
“How is your week going, Eleanor?”
“Just fine, Dr. Fischer. I was just trying to track down our order for more petri dishes. It was supposed to be here Tuesday, but we still haven’t seen it. I was worried that it was lost in shipment or back-ordered or something. The distributor told me that it didn’t make the weekend truck, but it was on the truck for tomorrow.” She fiddled with her pencil while she spoke.
“Thanks for tracking that down. I know Becky has been using a lot of plates lately. Everything else okay?” Addison felt like Eleanor seemed nervous and quiet again.
“Everything is…good.” She paused like she was going to say more and then shrugged.
“Okay. If you need anything, I’ll be in my office. I have to write a few exam questions now that I’m finished lecturing.”
Addison didn’t check in with Laina since she knew that her experiment wasn’t starting until after Anthony was finished. It bothered her that Eleanor seemed troubled again today and that Anthony had somehow messed up his reagents again. It seemed like he had turned over a new leaf in the last month, but now he was mixing up basic solutions wrong? She shook her head in frustration. Maybe he could get it straightened out and still save the experiment.
CHAPTER 48
Laina called Emmitt Thursday evening to update him with her experimental results. She had taken notes when observing Anthony’s experiments and knew what values she should get with the control conditions if her system was set up correctly. She knew going into this that Emmitt’s system might be more of a challenge than he was hoping.
“You couldn’t get the baseline readings established? Are you sure you had everything set up correctly?”
“Yes. I took very detailed notes and I watched him do this several times. It always starts out the same.”
“There has to be some way to manipulate it to get it in the right frame or level or whatever. Did you try altering the salt concentration?”
“I couldn’t do that today. All of the chemicals are in the main lab and I made up my solutions before I got started. Someone might have gotten suspicious if I went back and made a new solution mid-experiment. I’ll make a variety next week when I try again.”
“What did you tell Addison?”
“I didn’t tell her anything. Everyone was already gone by the time I cleaned up to leave. They asked me to lock up, which I guess means that they trust me.” She let out an awkward giggle.
“Well, what are you going to tell her? You know she’s going to look or ask when you see her tomorrow!”
“I’m just going to say that I must have had beginner’s luck last week and couldn’t patch a cell today.”
“I thought you said that Addison could look at your data since she’s the PI. She’ll see what you did and know that something is wrong.”
“I password protected my folder. Anthony told me that would keep everyone out.”
“That seems like a short-term fix. She’s going to ask why she can’t get into your file.”
“Maybe, maybe not. I think I can keep up the I’m still learning excuse for at least two or three weeks. It’s not unusual for people to struggle with this.”
“Someone might want to watch what you’re doing if you have that much trouble!”
“I’ve got this under control, Emmitt. Chill. This was just the first try. Things rarely work on the first try.”
“Except that in this case, they did work. They just don’t work the way we want them to. I need to send some data over to Apothecom soon to keep their interests.”
“I can send you some of Anthony’s. Just call it raw data. They won’t know. Or I could just adjust the numbers down to where they should be. I’ll fiddle with it when I go in tomorrow and see if I can make the numbers look passable for Apothecom.”
“How many lies are we going to compound onto this?” Emmitt threw up his hands in frustration.
“I don’t know, Emmitt. How many have you already told? And for how long?” She ended the call before he could scream at her.
Emmitt threw his stress ball at the wall and swore under his breath. He didn’t want any of his neighbors to call about excess noise or disturbing sounds, but he was really frustrated. He just needed one more thing to work. He did not need Laina to give him lip about it. He picked up the stress ball and started squeezing it again. Maybe he should go for a walk and try and brainstorm a solution to this roadblock. He didn’t know a lot about patch clamping, but surely there was a way to make adjustments. He grabbed his keys and his jacket and left the apartment. He knew there had to be a solution that allowed this to work.
CHAPTER 49
Over the weekend, Addison stewed about the dismal ending to the week. Anthony’s experiment was a waste. He never got the control run to work, so he couldn’t make any other measurements with that out of whack. Eleanor was acting strange again but said that she was fine. She had always been so cheerful and easy-going, so it was disturbing for her to be this recluse. Laina hadn’t had any luck patch clamping this week either, so that project was also at a stand-still. She wondered if there was a full moon that was wreaking havoc on her lab.
“You’ve been rather quiet all weekend. Want to talk about it?” Ryan asked as he sat down next her on the couch. Their kids had just left for youth group.
“No. I’m sorry. Ugh. I’m just frustrated. Is it that obvious?”
“You have seemed fairly distracted by your thoughts. I promise I won’t try to fix it or offer solutions if you want to talk. I will just listen. Scout’s honor.” He held up his fingers to his forehead in salute.
Addison laughed. “I hope I didn’t bring down the weekend for everyone. It was fun to go camping together again; it’s just been one of those weeks.”
Ryan waited to see if she’d continue.
“Eleanor is still not herself; it’s like she’s afraid to do something wrong, or I don’t know. She says that she’s fine, but her actions say that she is uncomfortable. Anthony completely wasted a week with his experiments, not to mention all the reagents. He either made up a solution incorrectly or messed up his transfection or forgot everything he’s learned up to this point. Laina couldn’t get a cell to patch this week either. She said that she broke a couple pipette tips and punctured some cells before she could get any measurements.”
“Surely something went right?”
Addison shrugged and looked at the ceiling. “I guess Laina finally got a new fob that lets her in the building without a special song and dance.”
“See? That’s something.”
“Juan has been really helpful getting Becky pointed in the right direction with her work too. I had to lecture almost every morning this week and that had me all out of sorts. Maybe I’m the one that’s out of whack and everyone else actually had a normal week.”
Ryan smiled. “I would never call you out of whack.”
She squeezed his hand. “Thank you. I’m sorry if I was a Debbie-downer all weekend. On top of all the stuff at work, I feel like our time with Joe is slipping away. I can’t believe he’ll really be in Colorado next year while we’re all still here.”
Ryan grabbed her hand. “I know. It feels like we really need to maximize our time with him, but he’s so busy. You weren’t a Debbie downer. We had a great weekend. I could just see the gears in your mind turning things over whenever we were relaxing in the camper or by the lake.”
“I’m not sure if I’m more frustrated that Anthony couldn’t get his act together again or more worried that something is wrong with Eleanor. I wish she would talk to me.”
“You could always call her. Invite her to lunch or something.”
“I don’t want to pressure her into it. I just want her to be herself or tell me why she feels like she can’t.”
“I guess you will just have to give her some time and space. She isn’t just your employee; she’s your friend too. I think she’ll come around.”
“Thanks. And thanks for getting me to talk. It’s much better than keeping it all bottled up inside. I wonder if Taylor is serious about looking at schools in the Denver area. Joe seemed really hopeful that they could stay together.” She sighed and gave him a hug. “Let’s see what we can scrounge up for dinner before those starving kids get back home.”
CHAPTER 50
On Monday, Addison sat down with Anthony to review his plan for that week’s experiment. She double-checked the solution recipes that he was using and had him write down all of the reagents that he would be combining. She could tell by the look on his face that he was frustrated by the entire process.
“I’m sorry to make you do this, Anthony. I can see that you are frustrated. I just don’t want you to waste another week like last week. It felt like you had really been making progress prior to that.”
“I know Dr. Fish. I can’t figure out what happened. I remade all of the solutions, but it didn’t change anything. Maybe I messed up the transfection somehow. I haven’t made a mistake like that in forever though.” He put his hands in his lap and looked at the floor.
Addison sighed. “I don’t want you to get discouraged. Maybe it was just an off week. I want you to double and triple-check everything this week. Be as absolutely methodical about every step as you can be. If it still happens, then we’ll back track and see what might have changed.”
“Okay. I will do my best. I hope it was just a fluke. Laina didn’t have any luck either?”
“She said that she couldn’t get a patch on a cell and that she broke too many pipettes trying to adjust the height with the microscope.”
“That’s too bad. I thought she would gain some confidence by working on her own this week. Hopefully, she isn’t too frustrated,” he said as he got up to leave.
“She seems pretty driven to get it figured out. I’m sure she’ll get it to work in the next week or two.” Addison mulled over the idea of asking Anthony about Eleanor, but decided it was too close to being lab gossip and kept it to herself. “Okay, well, good luck with this. I hope you realize that the end is in sight. You just need a few more good runs to pull this all together.”
Anthony nodded and left her office. She picked up the phone to call the lab. Maybe Ryan’s idea to invite Eleanor to lunch wasn’t such a bad one after all. It had been several months since they ate together.
“Hi, Eleanor. Are you busy for lunch tomorrow?”
“Tomorrow? I usually eat with the other techs on Tuesdays.”
“Oh, I didn’t realize. Is there a day this week that we could meet for lunch? My schedule is pretty free this week after being so busy last week.”
“Well, I guess I could do Friday,” Eleanor said slowly.
“Oh great! Friday it is. Is Santouits okay, or do you want to try some place new?”
“Santouits is fine.”
“Okay, I’ll swing by your desk around noon to walk over.”
“Okay. Did you need anything else? I need to get back to these orders.”
“No, sorry for interrupting. It’s been such a long time since we went for lunch and I thought we could catch up away from the lab. I’ll see you later.” She hung up and wondered why Eleanor seemed so nervous. They had worked together for almost two decades and she had never seen her act this way. She pulled out her phone and texted Ryan.
Eleanor is still not herself today. I took your suggestion. Lunch on Friday.
Hope she will talk to you.
Thanks. Love you.
Love you too.
Addison opened up her email to catch up on anything she had missed over the weekend. She downloaded the journal article to read later in the week and saw her reminder to talk to Becky about putting her committee together. She wanted her to have her first meeting in January to keep her on track with her project and the graduate school’s requirements. She knew that Becky still had two or three classes to complete before she would be in the lab basically full time. Addison sent Becky a quick email asking if they could meet one afternoon this week to put a plan together for her committee. If Anthony’s experiment went well this week, he could possibly get his committee to meet at the beginning of the semester as well. Maybe he would actually graduate this year.
CHAPTER 51
Over in the lab, Anthony was waiting for Juan to arrive. Juan had started coming back in earlier because he was helping Becky with her project. He wanted to talk about his experiment with the more experienced postdoc before he did the transfection on Tuesday. He was hoping that Juan had seen something similar to what Anthony observed with his cells last week and could explain how to fix it. Laina was in tissue culture and Becky hadn’t arrived yet either. It wasn’t that Anthony felt like he had something to hide, but he did have some apprehension nagging at him. He hoped Juan could set his mind at ease.
“Juan!” Anthony called out when he saw the young man enter the lab. “Have a minute?” He looked over to see if he’d caught Eleanor’s attention, but she seemed caught up with something on her computer and hadn’t even noticed Juan walk in. Juan set down his bag and walked over to Anthony’s desk.
“What’s up?”
“I had something weird happen with my experiment last week. Let me pull up the raw numbers on the computer here.” He clicked on his folder and found Thursday’s date. The folder had fourteen different files in it. He opened several to show Juan at once.
“Okay, this was my first try. See how the control level is already high?” He pulled up another graph. “I changed out the solution and it’s still high.”
“Huh. These are our regular cells?”
“Yes. See here? I tried putting in mexipres to see if it would lower the elevated response and it just killed the cell.”
“Did you try more than one dish of cells?”
“Yes, I tried three different dishes before I gave up. At that point, I was just pouring solutions down the drain for no reason.”
“What did Dr. Fish say? She has more experience with this than I do.”
Anthony squirmed. “I didn’t really show her all of it. I just told her that something was off.”
“Okay, let’s take a step back. Was there anything—anything that was different from other times you’ve set up an experiment like this? Different media in the dish during the transfection? Extra DNA, less DNA? New cells?”
“No. Everything was virtually identical to what I did the last three weeks.” He shrugged.
“I don’t know, man. It almost looks like a different channel. You didn’t add a potassium channel in there or something, did you?”
Anthony laughed. “I can barely keep up with one channel. Where would I find a different one?”
“I don’t know. I’m just brainstorming ideas. The response you saw to mexipres is especially weird. I have read about cell death with mexipres somewhere. I’ll try to see if I can find it for you before Becky gets here.”
“Thanks, man. How is all that biochemistry going?”
“She actually caught onto it really fast, so I’m just double-checking her plans for her at this point. I think she’s growing and purifying her proteins today and tomorrow for a big binding assay later in the week. Hopefully she’ll get some interesting results.”
“Well, I’m sure she’s going to be here any minute. I need to get my stuff out for tissue culture so I’m ready when Laina finishes up. I let her start first since I’m trying to trouble shoot things here still.”
Juan went back over to his desk while Anthony pulled his solutions out of the refrigerator. Anthony wondered if Juan still felt like Laina needed to be kept on a short leash. Maybe Juan had decided that she wasn’t trying to manipulate them after all.
CHAPTER 52
Laina waited to pull up her data until everyone else had gone to lunch. She didn’t want anyone to see what she was up to. She knew that Eleanor stuck to a strict thirty-minute lunch period so that she could start her commute before the rush hour traffic built up. She opened some of Anthony’s files from the experiments she had watched him do so that she could mimic his data as closely as possible without actually copying it. Emmitt said that they just needed something to convince Apothecom that they were headed in the right direction. That would get the ball rolling and allow them to start recruiting volunteers for the drug trial.
Laina had told Addison that she hadn’t gotten any data last week. Truthfully, she just hadn’t gotten anything that she could share with the lab. She didn’t know how familiar Addison was with the range of ion channels, but she didn’t want to raise any red flags with the collaborating PI. Laina flipped back and forth between Anthony’s data and her own. She pulled all of her data into a spreadsheet so that she could fine tune her numbers to look like his. On Thursday, she realized immediately that this wasn’t going to be the walk in the park that they had hoped it would be. Rather than just give up on the experiment, she still ran through all the conditions that she had planned. She wasn’t sure if the initial results would work for what Emmitt wanted, but having nothing was definitely not going to get them anywhere. It took a little finessing, but she eventually made her results look like Anthony’s. She saved the graphs and sent them over to Emmitt just as Eleanor came back into the lab. Laina smiled and looked back at her computer screen. She saved all of the doctored files into a file that she marked as hidden. Then she removed the password protection from her folder in case Addison got a wild hair to look at Laina’s trials from last week. She knew Emmitt was right when he suggested that password protecting her files would only arouse suspicion in the lab. The hidden file idea was a better option. She closed all of the files and grabbed her notebook to verify her transfection plans for next Tuesday.
