Rising Pressure, page 13
“Sheila can take you down to the lobby. I’ll see you in a few hours, Emmitt.”
An older woman with long dark hair stood up at the mention of her name. She smiled and opened the office door for them. They already knew the way to the main elevators but knew that they had to have an escort still.
“Right this way,” Sheila said, guiding them down the hallway. She rode the elevator down with them and watched as they exited the building before calling the elevator back to the ground floor.
“I think that went really well. What are your thoughts?”
“I’m anxious to get started so I can get back to California.”
“I understand that, but what do you think of the facility and the people?”
“They all seemed nice. I’m not sure if the small, blonde one trusts me though. She was pretty quiet and stand-offish.”
“Maybe she’s just shy.”
“I’m going to keep my eye on her. You never know with the quiet ones.”
“Are you okay for dinner? I know you haven’t been to the grocery store yet.”
“There is nothing that I want from a steak house. Gross. And for your information, I ordered my groceries to be delivered and they will be here at 8 a.m. tomorrow.”
“I know you’re a vegetarian, Laina.”
“I’ll see you tomorrow, Emmitt. Lab meeting. 9:30 a.m.” She turned and walked towards the entrance of her apartment building. Emmitt wondered if she was going to be angry about him not helping her unpack forever. He rolled his eyes and wished he didn’t need her help.
CHAPTER 34
Addison got home a little later than usual on Friday evening. Joe’s football game was out of town and while she hated to miss it, she was ready to just sit down and relax. Even though things couldn’t have gone more smoothly, it had still been a stressful two days. She grabbed her phone and dialed Ryan. The game was about to start and she wanted to touch base with him before it was too loud to hear.
“Hey, honey. How was your day?”
“It was long, but good. Anthony gave a great presentation of his work thus far for lab meeting and seemed to have won over both Laina and Emmitt. Becky even asked a couple of questions and wasn’t just a wallflower in front of the two of them, so that’s promising.”
“Did everything go okay with Laina getting a badge and a fob and everything?”
“It did. Eleanor walked her through it and made sure she had all the right forms before they went to the different offices. It was easy.”
“How were things with Juan and Anthony today? You said that things seemed tense between them yesterday.”
“I’m not sure. Juan and I worked on submitting his paper most of the afternoon, so I didn’t really see the two of them together today. Hopefully, it’s all blown over now. How are the kids?”
“We haven’t seen Joe yet. They should run onto the field any minute now. The cheerleaders are getting everything set up on the visitor side as we speak. Liv is sitting with her cross-country friends and I’ve got Martin next to me. His buddies didn’t come this time, so I thought it was better that we stick together with all the old people.”
Addison laughed. She could hear Martin groaning next to Ryan. “Okay, well, I will let you go. Cheer extra loud for me. I’m going to watch it online. Love you.”
“Love you too.” They ended the call and Addison wondered what she could have delivered for dinner. The Asian place up the street had recently started delivering. She liked their pot stickers and pad Thai bowl. Plus, while it wasn’t really fast food, they did put it together quickly, so she wouldn’t have to wait very long. She pulled up their app on her phone and placed an order for delivery. Then she grabbed a bottle of white wine from their fridge and poured herself a glass. She filled a water glass too and then turned on the TV while she waited for her food to arrive. She could stream the game over the internet and cast it onto the TV, which Martin had shown her how to do yesterday evening. She had written all the steps down because she was sure she wouldn’t remember otherwise. She texted Martin an image of the TV with the game getting ready to start to show off her new skill. He gave her a thumbs up in response.
Addison kicked off her heels and waited for the game to start. Joe had mentioned that there might be some college scouts at this game, so she hoped he wasn’t nervous about playing in front of them. She and Ryan had encouraged him to do his best and not think about the scouts, especially when he didn’t know for sure that they would be present. She yawned. Luckily, Liv’s cross-country meet was at home tomorrow and not until 10 a.m. She could sleep in a little bit before going biking with Martin and heading over to the golf course where they held the meet. Her team was getting to host the district meet this year, so it was their second home meet of the season. Next week was the regional meet and her coach thought that their team stood a good chance of qualifying for the state meet the following weekend. Addison yawned again. Hopefully she could get more sleep now that the collaboration was officially starting and virtually underway.
CHAPTER 35
By Monday morning, Addison was ready to be back at the office again. The weekend, while fun, had not been very restful between all of Saturday’s activities and then Sunday’s church activities. She had fallen asleep during the Falcon’s Sunday Night Football game against the Saints, which Joe and Ryan were appalled by. “Oh well,” she thought, “it’s not like my watching or not watching has any effect on the game. Not my fault they lost!”
After getting everything unloaded in her office and checking the mailroom, Addison swung by the lab to see who had arrived already. She knew Eleanor would already be there and probably Becky too. Juan had mentioned that he was going to be coming in a little later now that his paper had been submitted. She never knew with Anthony and didn’t know what to expect from Laina now that she had her own badge and fob. Eleanor was at her desk sorting invoices.
“Hi, Eleanor. How was your weekend?” Addison asked, wondering why Eleanor hadn’t even looked up when she entered the lab.
“Oh, it was fine. Quiet.” Eleanor said simply.
“Okay. Anything else I need to know?” Addison felt like she was getting in Eleanor’s way or distracting her, so she decided to forego their usual Monday morning chat where they recapped their weekend excursions.
“Oh, I think she is going to get it fixed, but Laina’s fob wouldn’t let her in the building this morning. Thankfully, she had her badge too, so another person let her in. She is over at the admin office seeing if they forgot to activate it or what.”
“That’s weird. Hopefully, it’s an easy fix and not too frustrating for her. I’ll be in my office if you need me.”
She turned to walk out of the lab and was almost run over by Laina herself.
“Well, that was an adventure, but I think it’s fixed now. They couldn’t figure out why it wouldn’t work. It worked fine with their system, so we took the elevator down and tried it on the door and at first, it still wouldn’t activate the lock. We came back up and he reset it and reactivated it. Then we tried it again downstairs and it seems to be working again. I guess this fob was made for this high-maintenance girl.” She smiled at Eleanor and Addison.
“Goodness. You would think they’d just give you a new one.” Addison said.
“He said that they were currently out of new fobs. Apparently, a lot of people have been losing theirs lately, so they placed an order for new ones. Of course, those are on backorder too. Hopefully this one will be okay for now. Is Anthony back yet?”
“I think he’s in the common lab still setting up some DNA cultures. I can walk you over there on my way back to my office.” Addison offered.
Laina nodded and followed Addison down the hall to the space they shared with three other labs, mostly for their biochemistry assays and growing cultures. Anthony was bent over a Petri dish and had three others stacked to the side. Addison said goodbye to Laina and went back to her office.
“Oh, hey, Laina. I didn’t hear you come in.”
“Probably because you’ve still got the Bunsen burner flaring full blast.” She said as she reached around him to turn off the gas.
“Oh, whoops. I was so focused on getting my sample spread out on this plate, I didn’t think about it. Thanks for shutting that off. Dr. Fish is always afraid that I’m going to burn down the lab or something.”
“Well, the Bunsen burner would be an easy way to do it.”
“Nobody’s perfect,” Anthony grumbled.
“A little testy today, huh?” Laina teased.
“Nah, just honest.” He smiled.
“What are you trying to grow?”
“I’m just getting some more of my two DNA samples to use in upcoming experiments as well as a couple that Becky uses too. I, uh, borrowed some from Becky a while back and need to replace those.”
“That’s very kind of you.” She looked at his ice bucket with the microfuge tubes in it. “Two of these tubes aren’t even labeled. How do you know what they are?”
“Labeling takes forever. I have my own system. Two lines for the channel, the alpha symbol for alpha9, you get the idea.” He pointed to the little symbols. “I write down the concentration in my lab notebook and aliquot them into multiple tubes of the same color to use. I get a new color when I run out and make note of that.”
“Yeah, that seems a lot easier than just writing a date and concentration on the tube in the first place, not to mention the actual name.” She rolled her eyes. “Looks like Becky labeled her tubes without issue.”
“Let me just stick these in the incubator and clean up. Then we can get started.”
Laina watched while Anthony cleaned up the space, including disconnecting the Bunsen burner from the gas line. He removed his gloves and scrubbed his hands clean at the sink.
“Have you done tissue culture work before?” He asked while he dried his hands.
“Not actively since grad school several years ago.”
“Well, stop me if I’m telling you something you already know. The most important thing is that you clean your hands well and then don’t touch your face at all while you’re under the hood. You would be surprised how fast bacterial contamination can spread.”
“That I remember. Remind me why the solutions are in this water bath?”
“The cells need to stay at about 37°C, so we warm all the solutions to that temperature too.”
Anthony walked her through sterilizing the surface with 70% ethanol and showed her which incubator they kept their cells in. Emmitt wanted Laina to use their cell line for the project, but they weren’t going to thaw those until Laina had gotten comfortable with patch clamping.
“Does anyone in the lab work on the weekends?”
“Umm, occasionally Eleanor or Dr. Fish will come in to feed the myocytes when we have those in the lab. Otherwise, everything is fairly self-sufficient. Sometimes, I’ll come in on a Sunday to set up cells for an experiment, but usually I stick to Monday through Friday.”
Anthony showed her how to tell if the cells were ready to be separated into more dishes, which was called splitting the cells. He demonstrated how to rinse a dish of cells with saline, and finally how to add back the media that provided sustenance for the cells.
“Will your cell line require special media? Or will you be able to use what we use? By the way, everyone has their own bottle of media and saline. That way if you only leave a little bit of solution in a bottle, you’re only hurting yourself.”
“Good to know. I will double-check with Emmitt, but I’m pretty sure that we just use the standard solutions.”
Anthony plated some cells in a few dishes and explained what sort of cell density you would want to have in the dish for patch clamping. He set up eight dishes for them to use later in the week in their practice experiment. Addison had encouraged him to set up his cells as he would for a normal experiment while he was teaching Laina. He could still get some data for himself while helping her learn the technique.
“Tomorrow, we’re going to do a transfection. Are you familiar with that?”
“Is that when you use the lipid solution to help get a DNA plasmid into your cells? I did them in grad school, but we haven’t needed to do them since we made our own cell line.”
“That’s pretty much the gist of it. I have a few different combinations of interactions that I’m wanting to test this week. Hopefully, we can get some more readings like last Thursday. If there is anything you’d like to test from your samples, just let me know. I’m going to set aside this extra dish of cells for you to use for practicing and learning. You won’t have any ready to experiment with until next week, though.”
“Sounds good. I don’t think I’m ready to start testing any of our reagents yet. We only shipped over a limited quantity and I don’t want to stress Emmitt out by running through it too quickly.”
“Okay, well, the next thing I’m going to do is finish analyzing my results from last week. You’re welcome to look over my shoulder, if you’d like to learn about that too.”
“Wonderful. I’m going to go check my email while you clean up here and then I’ll see you at your desk in the lab?”
“It’s a—” he coughed, “plan.” He blushed a bit as he almost said, it’s a date! Laina winked at him before exiting the tissue culture bay. He was glad that Juan wasn’t around to witness it. He would have just gloated about being right again.
He wiped down the tissue culture hood with alcohol again and put all of his solutions away. He secretly congratulated himself for really sticking to his new directive of being a better lab-mate. He hadn’t left anything out or lost any samples to his own delinquencies in over three weeks now. Maybe he had turned over a new leaf!
Laina was at her desk when he got back into the main lab room, but she heard him enter and closed down her browser window. She rolled her chair over to his desk while he booted up the computer.
“What’s really nice about our department is that we have a local network that we can use for file sharing, so what I save on the computer over there, I can still access over here, or any other computer that I go into. If I save it in the general “Fischer lab” folder, then anyone in our lab can access it, but if I save it in my own folder—” he paused while he hovered the mouse over a folder labeled awydrow, “then only I can access it.”
“Yeah, we have that too. I think the admin office was able to get me linked to this lab as well. I’m not sure if I have a folder yet or not.” She said, looking over his shoulder at the screen.
“Oh, you have to create that yourself. If I created it for you, you wouldn’t be able to open it. The only person who can access any of the files is Dr. Fish, but if you password protect your folder, she can’t get into it without help from the IT department. Anyway, let me just pull up my files from Thursday.” He clicked on his folder and dozens of other subfolders filled the screen.
“I label my experiments by date. So far, I’ve never done more than one experiment on the same day, so it works for me. Juan uses a more complicated labeling system.”
Laina nodded, “Well, I’ve seen how you label your microfuge tubes, so I’m not surprised to see that you use the simplest option to label your data files as well.”
“Some people are always critics. Anyway, here is my folder from Thursday.” He opened the file and six other files appeared. “Each time you record an experiment, it makes a new folder and labels it with the subfolder’s title, in my case the day’s date, and a number, starting with 001. You can change the file names later if you want, but this is how they are organized to begin.”
Anthony clicked open the first folder and showed Laina how they imported the data into a spreadsheet. Then he explained the calculations they ran to analyze the data from the experiment. He repeated these steps with each of the recordings from the previous Thursday and then created a graph to show Laina what he’d discovered.
“I have to repeat each condition with multiple cells on multiple days for any of the results to be statistically significant. It’s different from some of the biochemical assays in that you only need a total of three experiments for comparison. So, while you can get a lot of data really fast, you also have to repeat it more for it to be valid. I also have to show proof that we were expressing the various components by doing a Western blot. Thankfully, you only do that once. Wait, are you familiar with Western blots?”
“Yes, I assume you have your own antibodies to recognize each of your proteins, like the channel, your alpha9 protein, etcetera?” Laina asked.
“Yes, we have them tagged so that we can check for different proteins on the same gel. I’m going to make a gel and run it this afternoon if you need a refresher. I’m guessing you don’t do a lot of Western blots when you’re working in the crystallography field.”
“We do them on occasion. Once we know that we’ve isolated our protein and purified it, we don’t need to do them again, though. I’ll take you up on that offer. Thanks for all of your patience with me. I hope my learning curve isn’t too steep.”
“Happy to help. I want to have my own lab one day, so this is a great experience for my future too. I’ll catch up with you after lunch then.”
“Speaking of lunch, does everyone usually bring their own lunch? Do you eat together?” Laina asked.
“Eleanor usually eats with some of the other lab techs in the department, and Becky eats by herself in the student lounge. Juan and I will sometimes eat together. You’re welcome to join us; we usually eat right around noon.”
“I am used to buying my lunch every day, which it sounds like you two do not, so where should I pick something up?”
“Most people like the sandwich shop downstairs. It’s next door to our building. You can’t miss it.”
“Great. Thank you again.”
